Photoluminescent semiconductor materials

FIELD: measurement technology.

SUBSTANCE: porous-structured semiconductor materials are modified by recognition element and exposing to electromagnetic radiation carries out photoluminescence reaction. Recognition elements that can be chosen from bio-molecular, organic and non-organic components interact with target to be subject to analysis. As a result, the modulated photoluminescence reaction arises.

EFFECT: improved sensitivity.

31 cl, 13 dwg

 

The technical FIELD

This invention relates to materials for fotoaparati and identification with the target analysis, in particular materials having light-emitting properties, and more precisely to the photoluminescent semiconductor materials for fotoaparati and identification with the target analysis.

PRIOR art

Porous silicon (PSi) has been studied in detail for various semiconductor applications, after its opening in the late 1950's. Recently, PSi has demonstrated the ability to produce a strong luminescence in the visible region /1/, offering promising applications in devices(devices) optoelectronics silicon-based. Other porous semiconductor materials such as gallium arsenide, for example, were also reviewed /2/, but to a lesser extent.

Patents /3/ and /4/ describe a method for detecting chemicals at the termination of photoluminescence PSi and a corresponding device for the detection of organic solvents by using the photoluminescence PSi. To obtain the PSi silicon plate was subjected to electrochemical etching in a mixture of 50:50 hydrofluoric acid (HF) and ethanol. Upon irradiation PSi plate laser light source in the presence of organic compounds of the type of tetrahydrofuran (THF), diethylether, methylene chloride (MeCl2), Tolu is l, o-xylene, ethanol and methanol (Meon) characteristic fluorescent emission intensity of the PSi was significantly reduced (i.e., photoluminescent PSi reaction was suppressed). Also, Sailor /3, 4/ noticed ferrocene dissolved in toluene (i.e., cyclopentadienyl Fe (II)), resulted in complete loss of luminescence.

In General, Sailor /3, 4/ assumes that the degree of suppression of the photoluminescence (PL) of the reaction is determined by the dipole moment of the investigated compounds. Accordingly, it is possible to make the assumption that this evaluation method can help to assess differences in dipole moments between some organic compounds. However, Sailor /3, 4/ did not anticipate that this method can distinguish two or more part having a similar dipole moments, but significantly different in chemical composition. For example, in /3/ Sailor observed that MeCl2, Meon and THF had a relative suppression of luminescence, approximately 0.1 or less. Therefore, there is only a small difference in the dynamics of the PL characteristics produced by each of these compositions, which can be used to improve the characteristics of the corresponding chemical structure. Also, Sailor /3, 4/ notes the shift in wavelength of the reflected radiation (λapproximately 30 nm (1 nm=10-9m), from 670 nm to 630 nm, when PSi processed THF. He believed that the other is specified organic compounds also cause a reversible decrease in PL, but I had no idea that the shift X is similar to the shift λobserved(observed) for THF. However, on its face, it seems that Sailor /3, 4/ offers that shifts λ mostly similar in magnitude for all of these organic compounds. Accordingly, the 30 nm range defines several limited window for spectroscopic separation between the unknown compounds.

Lin and others describe biosensor based on the shifts of the wavelength at the edges of the Fabry-Perot reflection spectrum of stimulated emission thin film PSi visible range /5/. Optical thin film PSi, prepared by electrochemical etching in HF aqueous solution in ethyl 98% alcohol, are transparent 49%, which is sufficient to enhance the edges in the optical Fabry-Perot spectrum of the reflected radiation. Element recognition is fixed on the surface of the PSi film. Serial connection of the analyte with item recognition thus leads to a change in the refractive index of the PSi film and is detected as a corresponding shift in the interference pattern. The interference pattern is created by multiple reflections of white light between the surface of the partition solution/PSi and the silicon surface of the partition PSi/Substrate. Accurate and faithful interferometric spectrometrically strict requirements on the creation and maintenance of an almost perfectly parallel surfaces of the air /PSi, and PSi/ silicon substrate. Therefore, this technique is limited to applications where the environmental conditions (vibration, temperature changes, the composition of atmospheric gases) are strictly monitored.

Janshoff, etc. /6/ also describes PSi for biosensor applications using the shift in the interference pattern Fabry-Perot created by multiple reflections of white light at the boundaries of the air/PSi layer(interlayer) and PSi/BULK silicon surface section, as a way to detect types of intermolecular interactions in solution with bound ligands as receptors. Janshoff, etc. /6/ formulate that "a prerequisite for the use of porous silicon is the choice of the relevant parameters of the etching, since the optical and interferometric biosensor should also have an adjustable geometric shape then" (/6/ R. 12108). Thin silicon films were created porous by electrochemical etching, which helped to ensure that the pore radius ranging from 3 to 10 nm, a similar depth(height), and a cylindrical shape with absolute surface area of from about 0.1 to 0.15 m2for samples, protravlivaya 1 cm2of silicon. Excessive porosity is not suitable for biosensors note Janshoff, etc. /6/.

Since interferometric methods are based on the specific material is " a phenomenon namely, the light reflected from the boundaries of two different planes, producing an interference pattern, a biosensor system that relies on shifts in the interference pattern to detect the presence of the analyte, usually very sensitive to vibration, temperature and changes in atmospheric pressure. In addition, the reflective surface of the film or plate, that is, the air/PSi and PSi/silicon substrate boundary surfaces must be parallel, or may occur an undesirable shift in the interference pattern. Typically, the reflective surface of the film PSi should be parallel to 25 a (2.5 nm), which imposes high requirements on the level of production of PSi plate or film. Finally, for the optimal characteristics of the light directed onto the film PSi or plate must be perpendicular reflecting surfaces. Accordingly, the pores that are not directly perpendicular reflecting surfaces, affect the interference pattern film PSi or plates and, therefore, adversely affect the results obtained from such a biosensor.

It is therefore desirable to have a material adapted to detect the analyzed compounds, which could use the photoluminescence properties of porous semiconductor materials. In addition, that the second material could be increased sensitivity to detect low concentrations of the analyzed compounds.

The INVENTION

In accordance with this invention photoluminescent semiconductor material is a composition of modified semiconductor composed of at least one of a semiconductor material having a porous texture, and at least one element of recognition, and when covering the specified composition with electromagnetic radiation, at least one wavelength in the range from 100 nm to 1000 nm, the composition produces at least one first luminescent reaction in the form of electromagnetic radiation, at least in the range from 200 nm to 800 nm.

In accordance with another aspect of the present invention proposes a method of analysis of a substance in the luminescent reaction, including the irradiation of the composition of the modified semiconductor comprising at least one semiconductor material with a porous structure and at least one element recognition, electromagnetic radiation, at least one wavelength in the range from 100 nm to 1000 nm, obtaining at least one first luminescent reaction in the form of electromagnetic radiation from the said semiconductor composition, dimension, at least, intensity, or wavelength, at least one first the fluorescent reactions is.

BRIEF DESCRIPTION of DRAWINGS

See drawings for explaining the embodiments of the proposed invention.

Figure 1 - schematic representation of a variant of the device design that can be used to detect the photoluminescence from the photoluminescent devices according to this invention;

Figure 2 is a detailed electronic micrograph of porous silicon particles produced in Example 1, 300-fold increase;

Figure 3 is a detailed electronic micrograph of porous silicon particles produced in Example 1, 800-fold increase;

4 is a detailed electronic micrograph of porous silicon particles produced according to Example 2, 800-fold increase;

5 is a detailed electronic micrograph of porous silicon particles produced according to Example 2, at 5000-fold magnification;

6 - EPI-fluorescence micrograph of 200-fold magnification, showing the interaction of the antigen with the PSi material fragment modified with antibody discussed in Example 10;

Fig.7 is discussed in Example 13, a graphical comparison of the hydrolysis of acetylcholine to enzyme control in comparison with modification PSi enzyme;

Fig is discussed in Example 16 fluorescent EPI-fluorescence micrograph of 200-fold increase, which illustrates the output of the antibodies is PSi, not treated with the buffer aminouksusnoy acid;

Fig.9 is discussed in Example 16 EPI-fluorescence micrograph of 200-fold increase, which demonstrated a decrease in the fixing antibodies PSi, treated with buffer aminouksusnoy acid;

Figure 10 is discussed in Example 16 EPI-fluorescence micrograph of 200-fold magnification, showing the binding activity of the antibody figure 9, does not adversely effect the buffer aminouksusnoy acid;

11 is a graphical representation of the intensity of the photoluminescence of porous silicon material fragments discussed in Example 17;

Fig is a graphical representation of the intensity of the photoluminescence of porous silicon material fragments with attached item recognition (see Example 17).

Fig is a graphical representation of the intensity of the photoluminescence of porous silicon material fragments, in the analysis of substances in contact with the element of recognition discussed (smpnameres 17).

A DETAILED DESCRIPTION of the BEST IMPLEMENTATIONS

In accordance with this invention the porous semiconductor (PSc) has a photoluminescent (PL) properties and can be used in a number of applications for detection and identification, including, without limitation, biotechnology, type biosensoric, medical, control the he environment, for industrial use and protection (for example, to detect the damaging chemical and biological substances), genetic diagnostics and other molecular/cell biological applications of type selection/sorting cells, microstructural analysis of cells, etc. In accordance with the invention PSc materials have, at least, the PSc component with a porous texture, the modified at least one element of recognition. In the PSc material introduced an element of recognition to interact with the analyzed objective (i.e., the substance to be detected). Element recognition is usually, but not always modifies PL reaction PSc material, produces a shift of wavelength λ and/or changes in the intensity of PL compared with the reaction of the PSc material that does not contain the element of recognition. PSc modified such recognition elements ("PSc/RE") can interact with analyzed in order, shifting the wavelength and/or by changing the intensity of the PL, and this PSc/RE reaction will be referred to simply as the modulation PL reaction or PL modulation for short.

PSc material.

Preferably, as the PSc material was used silicon. However, the PSc material may include any semiconductor material composition, which has the properties of photoluminescence in porous the structure. Such semiconductor materials and compositions may include, without limitation, cadmium, copper oxide, germanium, gallium, gallium arsenide, selenium, silicon, silicon carbide, silicon dioxide, silicon phosphide of gallium, and combinations thereof. Selected semiconductor material compositions may also include alloying substance, including, for example, without limitation, erbium, boron, Fosforit, copper, phosphors several lanthanides, including ytterbium, holmium and thulium, and combinations thereof. Also, the selected composition of semiconductor materials can be to have a different composition, including, for example, without limitation, halogen type, bromine, capable of changing the wavelength of the radiation Bressers and other /7/. For ease of discussion, it should be understood that the term "PSc" includes, without limitation, any semiconductor composition material, of the type mentioned above for illustrative purposes.

The PSc Structure

The term "porous" or "porous texture" is used by us to characterize any disturbance, type of recesses, protrusions, or combinations thereof, in or on the semiconductor material that forms the complete surface area of the semiconductor material. Examples of the recesses include, without limitation, pores, ripples on the surface of the plating, shell craters, trenches, grooves, and combinations thereof. Examples include protrusions which, without limitation, combs (jumpers, edges, welts, bumps), and combinations thereof. For example, without limitation, such disturbances can be in the form of ordered cell porous structure, comprising a cylindrical or polygonal shape of the pores or more random porous structure, which is similar to the coral.

The shape and geometry of the perturbation of a semiconductor material can be varied. The recesses can vary from regularly-recurring shapes and configurations to irregularly-specific forms and configurations and combinations of these. Also, the protrusions may vary from regularly-recurring shapes and configurations to irregularly-defined shapes and configurations. Regularly, certain forms include without limitation, circular, semi-circular, ellipsoidal, paleolimnology, polygonal, square, rectangular, triangular, rhombic, trapezoidal shape. Irregular forms include, without limitation, mixtures of the above forms of disturbance. Also, for example, without limitation, 3-dimensional (three-dimensional) geometry perturbations may change from regularly-defined three-dimensional structures to irregular three-dimensional configurations. Regularly-defined three-dimensional configurations include, without limitation, cylindrical, conical, cubic, parallelepiped, mnogogranen the ka, rhombohedral, elliptical, spiral, spherical and pyramidal forms. Irregularly-specific three-dimensional configuration include, without limitation, a combination of the above forms. In any case, such disturbances in the semiconductor material result in increased area of surface PSc material.

Full geometric shape PSc material may be in the form of a film or plate, or to have a three-dimensional structure. Under the "three-Dimensional structure" we mean such a geometry PSc material in which the material itself or in combination with the support of non-PSc material may be a variation regularly-defined geometric shapes and irregular-specific forms and their combinations. Examples of regularly-specific forms include, without limitation, spheres, half-spheres, ellipsoids, problemcode, cylinders, ovate, rods, disks, funnel, cubes, parallelepipeds, polyhedra, rhombohedrons and pyramids. Examples of irregularly-specific forms include, without limitation, a combination of the above PSc forms of material. For ease of discussion, the reference to "the PSc structure" means, without limitation, the PSc material, which is in the form of a film or plate, or which has at least one of the types of geometric shapes and porous texture described above for illustrative purposes.

Fig. 2 through 5 illustrate only a few of the types of surface perturbations that characterize the PSc materials in accordance with this invention. Obviously there are many technologies and methods that can provide a wide variety of perturbations on the PSc. As will be discussed in further detail below, regardless of the specific form or geometry of such disturbances, these disturbances increase the PL intensity of the radiation produced by the PSc material. Thus, without reference to appropriate theory it is clear that the porous perturbation on the surface contribute to the increase in PL intensity.

In accordance with a preferred variant of the constructive embodiment of the invention, the PSc material has a porous texture on one or more surfaces and/or throughout its structure.

The actual shape and/or size structure of the PSc depend on the specific application in which the structure of the PSc is used. There are some applications in which the flat structure of the PSc is desirable and others, where a three-dimensional structure of the PSc is more preferable. When the three-dimensional structure of the average diameter or other largest average size should preferably be in the range from 100 nm to 1 mm, Most preferred are the three-dimensional structure of the PSc, the average diameter or other largest average times the EP which is in the range from approximately 100 nm to approximately 500 microns. The most preferred three-dimensional structure of the PSc have an average diameter or other largest average size in the range from about 1 micron to about 500 microns. In addition, in cases where the structure of the PSc - dimensional structure of the PSc, it should be adapted to a number of device configurations of fotoaparati, and its landing size should be consistent with the appropriate size of such devices fotoaparati. For example, when used in capillary diameter of three-dimensional structures of the PSc should be in the range of from about 5 microns to about 15 microns.

The structure of the PSc of the present invention, subjected to illumination by electromagnetic radiation of a wavelength in the range from approximately 100 nm to approximately 1000 nm, sufficient power, produce fluorescent radiation in the range from about 200 nm to about 800 nm, preferably in the range of from about 350 nm to about 700 nm, and more preferably in the range of from about 450 nm to about 650 nm.

The structure of the PSc in this invention can be formed in various ways that are obvious to a skilled in this technology specialists. Flat or similar plate of semiconductor material is commercially available, for example from /8/. Also, for example, not having the silicon spheres can be produced by the process, described in /9/. Further, flat or similar plate of semiconductor material can be mechanically fragmented to produce thin material point irregularly-defined configurations. In any case, regardless of the procedure selected for production of a semiconductor material, such material may be made porous in accordance with a number of known technologies, such as those discussed below for illustrative purposes.

After dense semiconductor structure created it is made porous. Desirable porous texture can be obtained in a variety of known methods. For example, without limitation, the porous structure can be produced by epitaxial deposition or lithography /10/, chemical etching /11/, anodized in HF solutions /10/, spark erosion /12/laser ablation /13/, milling ion beam /14/ and managed by annealing and etching /15/. Also, qualified specialists it is clear that the technology used for the formation of the porous structure to a mostly flat semiconductor material, may require some changes. For example, a three-dimensional semiconductor structures may require suspension in an inert gaseous or liquid environment, so that all surfaces come in contact with isomerisations source, an etching solution or other means of forming the desired structure of the PSc. Also, the time given for etching, may also be much shorter than the three-dimensional structures of the PSc.

The structure of the PSc can be formed with a channel for a heat transfer fluid. Examples of relevant core materials include, without limitation, glass, plastic, ceramics, zeolites, metals, various semiconductor materials, and combinations thereof. The material of the active zone can be selected, for example, from the condition of attaining the desired density for a specific application. This kernel (the channel for the heat transfer fluid)is likely to be more acceptable in the case when the patterns have a larger diameter. In the case of a single coating of semiconductor preferably, this coating should be thick enough so that it could form the desired porous texture.

Floor base PSc material can be performed not from the porous material by a number of methods. For example, dense coating kremmidiotis or silicon carbide can be obtained by controlled oxidation (/16/ or high-temperature pyrolysis methods /17/. After that, the semiconductor material with the coating can be converted to a porous state. Alternatively, the structure of the PSc can be formed simultaneously coated with the eating of the coating on the surface of the active zone material. Also, it may be desirable application of multiple layers of different types on the surface of the active zone material.

Item Recognition

In the present invention, the structure of the PSc modified at least one element of recognition and, preferably, many elements of recognition. For simplicity recommendations, the structure of the PSc, the modified at least one element of recognition hereinafter referred to as "PSc/RE". The term "Modified" is used in the sense that the element of recognition associated with the PSc structure so that PL reaction PSc/RE is modulated when the analyzed object interacts with the element(s) recognition PSc/RE. One example of such modifications is a covalent bond item recognition with the structure of the PSc. However, direct communication or material may be absent. Maybe the closest Association item recognition with the PSc structure sufficient to maintain or electronic energy transfer between item recognition and structure of the PSc. Typically, but not necessarily always, PSc/RE shows increased PL reaction relative to the PL of the reaction for the structure of the PSc before modification with item recognition

In General, the recognition elements may be organic, inorganic, biomolecular to the components and their combinations.

Preferably, as elements of recognition performed biomolecular components. Examples of biomolecular components that can be used as elements of recognition, without limitation, are natural or synthetic proteins, nucleic acids, oligonucleotides, lectins, carbohydrates, glycoproteins and lipids that interact with the subject of the analysis. Proteins are preferred as an element of recognition. Examples of suitable proteins, without limitation, are the immunoglobulin type polyclonal and monoclonal serum, and enzymes. A privileged position among the elements of recognition is occupied by proteins such as immunoglobulins and enzymes. Examples of relevant nucleic acids are simple chain molecules DNA and double-chain molecules ONA. The element of recognition may include attached redox component. Examples of redox components include, without limitation, transition metals and combinations thereof, and co-enzymes-type NAD (H) or NADP (H). Redox components can be of type e donor or acceptor, contributing to the change in PL intensity in the interaction of the analyzed item by item recognition.

Examples of the inorganic component is in, which can be used as recognition elements, without limitation, doped or undoped crystalline inorganic compounds, such as linear carbon modifications, including carbine and diamond crystals.

As examples of organic components, which can be used as recognition elements, without restrictions can be named polymers with internal conduction ("ICP") type polyaniline and a polymer electrolyte type, polyethylene oxide, mainly useful doped with lithium.

The change in the structure of the PSc item recognition can be enhanced by the use of surfactants that reduce the surface tension of the solution, with the element of recognition. Such substances include, without limitation, alcohols and detergents in concentrations sufficient to reduce the surface tension of the solution without adverse effects on the structure of item recognition, the performance of the cell recognition or PSc/RE.

Analyzed the subject

As a result of interaction of the analyzed object with PSc/RE is the modulation of the PL response. We assume that at the interface of the analyzed object communicates with item recognition, modulates PL reaction PSc/RE. Examples of such interactions include, without limiting the tion, the covalent bond, the formation of hydrogen bond, the strength of the van der Waals forces, and srodstvenny communication. However, direct communication or material may not be required. Sufficient Association of the analyzed object with PSc/RE to maintain electronic or energy transfer between the analyzed object and PSc/RE.

Analyzed the subject may be organic, inorganic or biomolecular structure or composition. Examples of analyzed subjects include, without limitation:

(1) antigenic compositions that comprise antibodies, including, for example, without limitation, toxins, metabolic regulators (stabilizers), microorganisms such as bacteria, viruses, yeast, fungi (fungi) and microbial spores of animal and vegetable origin, elements, fabric;

(2) the specific materials such as enzymes, including, for example, without limitation, metabolites, specific substances, pesticides, insecticides;

(3) the complementary oligonucleotide sequence;

(4) simple chain molecules of DNA and RNA;

(5) the ligands of hormonal receptors and lectins.

In the interaction of the analyzed object with a certain element of recognition on the structure of the PSc is the change in the PL response of the structure PSc relative to the PL response of the structure PSc/RE without the analyzed subject. Preferably the, to PL reaction was increased relative to the base PL reaction PSc/RE. Modulation PL reaction can be identified in real-time or post-processing of results.

Stabilization/Activation Patterns PSc

In order to maintain a stable and effective response patterns PSc with item recognition, the surface structure of the PSc first stabilized to prevent uncontrolled oxidation.

One variant of this method of stabilization is the use of oxidation, for example, without limitation, thermal oxidation /18, 19, 20, other - oxidation process in the atmosphere of ozone. These processes produce reactive hydroxyl groups. Chemical oxidation can be produced, for example, using peroxide, dimethyl sulfoxide (DMSO) or powder iodine.

Covalent Bond

As noted above, in one of the preferred embodiments of the present invention, the structure of the PSc can be modified element recognition covalent bond. For example, the element of recognition may be covalently linked to the structure of the PSc via one or more linker. The linker may provide functional move groups of chemicals groups on the structure of the PSc for access to the item recognition. Also, among other functions, the linker may reduce spatial sweaty the material, that eliminates difficulties in solving the problem of identifying the subject of the analysis and its interaction with the element of recognition and/or PSc/RE.

Preferably, the element recognition covalently linked to the linker so that the element of recognition and/or PSc/RE can interact with analyze the item with maximum efficiency. For example, some of biomolecular components of the type of antibody, conjugated through their sulfhydryl groups, show good results when interacting with the subject of the analysis. However, when such antibodies attached via amine groups, they can reduce this potential and, thus, to facilitate interaction with the subject of analysis.

The main layout

In another preferential embodiment of the invention the main linker attached to the hydroxyl groups resulting from the oxidation of PSc. One example of the main linker - substituted silane. An example of a suitable substituted silane, without limitation, glycidoxypropyltrimethoxysilane. This is the main linker provides a direct link between the hydroxyl groups of the oxidized structure of the PSc and the amine group of item recognition. Other appropriate chief linker - hydrosilicate alkenes and alkynes.

Other linker, which is reactive to hydroxyl the m groups of the oxidized PSc and the amine groups of item recognition, it is quite obvious for this technology. It is clear that the linker can be selected and thus to react with a functional group, other than the amine group of item recognition. Other functional groups item recognition presents sulfhydryl, carbohydrate or carboxyl groups. Also understood that other linker, which is reactive to digitoxigenin groups (other PSc methods of stabilization) and functional groups selected elements of recognition, can be selected to communicate with the PSc structure.

The main and Secondary linker

In another preferential embodiment, the main layout is associated with hydroxyl groups resulting from the oxidation of PSc and then the secondary link linker attached to the main linker. One example of the main linker that can be used in combination with a secondary connection between the linker - substituted silane. An example of such corresponding substituted silane, without limitation, aminopropyltriethoxysilane. Examples of the corresponding secondary communication linker without limitation, Homo - and/or heterobifunctional cross-linking agents and hydrosilicate alkenes and alkynes.

Because the secondary communication linker typically cannot communicate directly with the PSc main compone the expert provides direct interaction between the structure of the PSc and the reactive group of the secondary communication linker. Accordingly, the main and secondary linkers provide indirect interaction between the PSc and the element of recognition.

Also, using the connection of the secondary linker with the main linker, provided even more long intermediate compounds in comparison with one main linker. Therefore, among other functions, mixed linker consisting of the main and secondary, can solve the problem of reducing the spatial potential resulting from unusual volume of subject analysis, and contributes to a more successful search item interaction recognition and/or PSc/RE. Secondary link Builder may also provide greater flexibility in the choice of the functional group that is reactive, for example, amines, sulfhydryl, carbohydrate or carboxyl groups of the recognition elements.

Homobifunctional cross-linking agents have two such reactive groups. For example, homobifunctional crosslinking agent may have a first reactive group, which can interact with the amine group of the main linker and the second reactive group, which can interact with the amine group of item recognition. Examples homobifunctional cross-linking agents, without limitation: the aldehyde of glutarate, disuccinimidyl suberate, E. what about the sulfonated analogue and again (sulfosuccinimidyl) suberate.

Heterobifunctional cross-linking agents have two different reactive groups. For example, heterobifunctional crosslinking agent may have a first reactive group, which can interact with the amine group of the main linker and the second reactive group, which can interact with sulfhydryl group of item recognition. Examples heterobifunctional cross-linking agents - Succinimidyl 4-(N-maleic aminomethyl)-cyclohexane-1 - carboxylate, and its sulfonated analogue - 4-(4-N-maleic AMINOPHENYL) hydrazide-Hcl butyric acid, and 4-(p-azidoaniline) butylamine.

The main layout, which is reactive to the hydroxyl groups of the PSc and functional groups selected elements of recognition, it is quite obvious, on the basis of the described technology, and it is also clear that other linker, which is reactive to digitoxigenin groups (other PSc methods of stabilization) and functional groups selected elements of recognition, can be selected to communicate with the PSc structures.

PSc/RE Interaction with the Subject of analysis

The use of item recognition, which has a specific affinity for the subject of analysis is highly desirable, because it reduces the likelihood that a foreign object that is included in the analyzed composition, in aimogasta with item recognition. Similarly, the use of item recognition, which facilitates the characteristic modulation in PL reaction PSc/RE, when the analyzed object interacts with the PSc/RE, will reduce the likelihood of "false" modulations produced by extraneous compounds. Accordingly, the impact, if any, impurities included in the analyzed compositions PL reaction can be significantly reduced.

Similarly, in an advantageous embodiment of the invention, where the element of recognition has a specific affinity for the item analysis item analysis mainly interacts with the element of recognition rather than with the structure of the PSc.

The structure of the PSc of the present invention should contact the subject of analysis in many different applications, examples of which are given below. In some applications, it may be desirable to place the PSc/RE in the corresponding transport in order to increase the interaction PSc/RE to the subject of analysis. Using the contacting means, evident in the proposed technology, vehicle/PSc/RE mix may enhance the degree of contact between the unknown composition or body, regardless of whether plant or animal, they contain the subject of the analysis or not. Such container may include means for reducing the surface tension chosen to replace the first device. You can increase its hydrophilicity, or to increase its hydrophobicity, as appropriate. Such means include, without limitation, alcohols, detergents and organic solvents in a concentration sufficient to achieve the desired effect without adverse effects on the efficiency of the PSc/RE.

Processing linker

The linker can be processed for

(a) amplification of a preferred interaction between him and the item recognition, and/or

(b) slowing down the interaction between unrelated linker (i.e., the linker is not associated with item recognition, but paired with the PSc) and compositions without the analyzed object and/or purpose of analysis.

Preferably, the interaction between the linker and the item recognition has been increased that will provide a stronger Association between item recognition and linker and/or Orient the element of recognition for the best interaction with the subject of analysis.

The interaction between unrelated linker and non-target compounds and/or the analyzed objects may slow down when you lock the reactive groups of unrelated linker, and/or accession to the reactive groups of unrelated linker components that have an affinity for structure, not present in the pre the meta analysis.

An unlimited number of examples of such compositions for the treatment of the linker are discussed in more detail below, they are divided into:

(1) with immunoglobulin binding protein),

(2) Biotin reactive agent and

(3) blocking solutions, such as buffer solutions Amin.

Treatment with immunoglobulin for anchoring of the protein.

In one implementation options of the linker attached to the structure of the PSc can be processed protein-associated immunoglobulin ("IgBP") (e.g., Protein a, Protein G or Protein L). lg has a specific affinity for a particular part of the antibody is known as the Fc domain. Therefore, since all antibodies have the Fc domain, element detection antibodies will preferably interact with IgBP. Therefore, when the element detection antibodies come in contact with IgBP processed, the linker is attached to the structure of the PSc, the Fc domain of an element of recognition of the antibody interacts with IgBP - processed by the linker.

Because IgBP has a specific affinity for the Fc domain of antibodies, IgBP processing linker reduces the chance of joining foreign compounds and analyzed subject to the linker, because such compositions and analyzed the items of course have no Fc domain. Another advantage of the IgBP processing linker is that the elements of u is syvania antibodies are properly oriented for interaction with antigenic analysis. Namely, the Fc domain of element detection antibody is attached to IgBP, while the Fab domains of the antibodies remain free to antigenic interaction.

Processing biotinyl chemically active agent

In another embodiment, the implementation of the linker attached to the structure of the PSc can be processed biotinyl chemically active agent ("BRA"), which has an affinity to Biotin. You can bring an unlimited number of examples bilinovich chemically active agents - streptavidin, neutravidin and avidin. In this case, the desired item recognition treated with Biotin to item recognition was consistent. Accordingly, when treated with Biotin item recognition comes in contact with the TREATED BRA linker joining the PSc structure, the component of the Biotin on the element of recognition is selectively interact with the BRA.

Because the BRA has an affinity only for Biotin treatment BRA linker reduces the probability of interaction of foreign compounds and analyzed objects without Biotin, with the linker. Another advantage BRA processing linker is a stronger relationship between the elements of recognition and a linker attached to the structure of the PSc. In addition, treated with Biotin element recognition is able to interact with predmeta the analysis.

A binder solution for processing

In the following implementation, PSc/RE can be processed with a binder solution to block communication of the reactive groups unreacted linker with strangers compositions and subject analysis. Solutions block include, for example, buffers amine, type a buffered solution aminouksusnoy acid. Any reactive group unreacted amine linker thus blocked amine groups of the blocking solution. However, the solution blocking does not block the receptor or acceptor element scope of recognition, which remain sensitive to interact with the subject of analysis.

PL Detection

The design variant of the device 10, which can be used to detect PL, schematically shown in figure 1. Sample 19 is placed on a typical tripod 20. The light in the predetermined wavelength is directed at the sample from an argon ion laser 12. Narrow band filter 14 reduces any noise radiation caused by the gas fluorescence in the chamber of the laser 12. The filter 14 absorbs radiation of all wavelengths, except for the radiation having the selected wavelength. The modulator 16 is used to suppress any constant noise, caused by any peripheral radiation. The modulator 16 provides modulation of the laser radiation is some frequency, which simultaneously separates the PL signal from the noise in the amplifier 42. After the modulator 16, the light passed through the lens 17 to the sample 19. The radiation scattered by the sample 19 is directed through lens 18 and a broadband filter 22 to the entrance slit 32 and mirror 34 in the monochromator 30. Broadband filter 22 absorbs radiation of wavelengths outside the specified range and thus reduces the laser beam reflected from the optical elements of the device 10.

Rotating diffraction grating 36, located inside the monochromator 30, takes into account the angular separation of the various wavelengths λ1that λ2that λ3and λ4from the scattered spectrum of the beam to the mirror 35. Rotation of the grating 36, controlled by the computer 40, allows you to select different spectral components emerging from the exit slit 33 of the monochromator 30 radiation. The intensity of the spectral components λ1that λ2that λ3and λ4registered by the photodiode 38. The signals of the photodiode 38 are amplified in amplifier 42 and fed to the computer 40. All measurement processes are controlled by the computer 40.

Quantitative determination of the concentration of the subject of analysis can be calculated using the calibration curve of the PL intensity for known concentrations of the analyzed object or using different concentrations of the element is raspoznavaniya.

Application

Patterns PSc/RE can be used by themselves or in combination with a wide variety of devices commonly used in a variety of applications, including, without limitation, biotechnology type biosensoric, for medicine, environmental monitoring, for industrial use and protection (for example, to detect the damaging chemical and biological substances), genetic diagnostics and other molecular/cell biological applications of type selection/sorting cells, microstructural analysis of cells, etc.

For example, there is a great need for precision devices, screening for genetic technologies and diagnostics. In the invention, individual parts of structures PSc/RE having different recognition elements can be prepared and then configured and/or harmoniously coordinated with bicrystalline or microcapillary multiple configurations, discussed in more detail below. The analyzed samples may come into contact with fixed or suspended structures PSc/RE in these configurations and interaction with the subject of analysis can be provided as discussed above. Very valuable that the three-dimensional structure of the PSc can be used with such technical means, such as automated valves microvolume, analytic found what whitelam and/or microprojectile identifiers.

In one embodiment, the PSc/RE patterns are effectively used for the detection of a photon of biocrystal, and the number of recognition elements can be placed in different areas of the same device. To date, the practical problem of multiple locations, but the various elements of recognition on a small semiconductor chips has not yet been satisfactorily solved as a result of:

1) insufficient signal intensity for detection of interaction with the subject of analysis and

2) difficulties accommodate the many different elements of recognition within a limited area. Small three-dimensional structure of the PSc, described above, are most suitable for this type of application. Sample containing an unknown composition, analyzed according to one of the many compounds can thus be identified by contacting biocrystals formed by the elements of recognition from a number of prospective structures. When biocrystal produces modulated PL reaction, the composition is identified by the element of recognition used in this specific area.

PSc/RE patterns can also be placed in microprojector, each individual region which contains exactly the set number of patterns PSc/RE and each individual region has the specific settings for different analyzed items or that the same has analyzed the subject, such that the interaction of the subject of analysis leads to the PL modulation, for example with unique characteristics (recognition samples).

In another embodiment, the implementation of the PSc/RE patterns can be used as packing in a microcapillary column, which can determine the analyte contained in the pumpable fluid. Microcapillary columns must be optically transparent in order to detect the PL modulation after interaction with the subject of the analysis. Parallel or serial arrangement microcapillary columns can be used to detect and identify one or more of the analyzed substances. In another configuration microcapillary columns may also include fiber optic elements, catching and detecting such PL modulation.

In one embodiment, the analyzed object can be located in or on the body or body part. The "body" referred to microbial cells (cells), animal and plant cells and tissues, microbial spores, viruses and the like, for Example, without limitation, analyzed the subject can be a bacterium, virus or microbial dispute. Examples of such analyzed subject include spores of the anthrax Bacillus and E. type E. Bacillus //0157 (factor disease hamburger). PSc/RE structures is, which bear resemblance to one of the analyzed objects come into contact with a sample, suspected in the presence of such of the analyzed subject, or a sample test for the presence of such of the analyzed subject one or more recognition elements attach themselves to the wall of the cage or floor through external receptors of the cells. It is very useful that these patterns PSc have a diameter or largest size from about 100 nm to 1 micron.

PSc/RE can be entered or pasted into the body for intracellular detection of the object of analysis.

Thus, an unlimited number of examples of various embodiments of the invention may be made for purposes of illustration.

EXAMPLES

Example 1 - Production of Psi particles

Small silica particles were produced by mechanical fragmentation of flat silicon n-type, obtained from Silicon Mines (Santa Clara, CA, USA). Particles were produced by mechanical fragmentation, using a mortar and pestle to produce granular particles of irregular shape. On this procedure, there were obtained particles in a wide range of sizes. Particles with a diameter or other largest size, between 30 and 1000 μm were selected mechanical sifting through multiple cellular is embrane. The particles were analysed using brüning-Emmy-Tolera (BET).

The BET analysis was done using micrometric device /18/. Empty sample tubes were weighed and then re-weighed after placing them in the sample. Samples were degassed by heating to 180°and fast moving them in a vacuum. After degassing the samples were re-weighted. Samples were also analyzed and were weighed again after the analysis. This last recorded weight was used for calculations in the analysis. The coefficient of volume absorption was taken from 0.05 to 0.3 relative units.

The surface area of the particles calculated based on the BET analysis part of the surface with 5 points, ranged from 0,1724 to 0,0016 m2/g with correlation coefficient 0,999871.

The silicon particles have been made porous, with a random distribution of pores, using the method of chemical etching. The silicon particles were suspended in 70% nitric acid solution, 50% fluoride-hydrogen acid and water in the ratio 1:4:1 for 60 seconds at room temperature. The reaction produced hydrogen in the form of gas and caused strong mixing of the solution, such that no additional mixing was not required to maintain the particles in suspension. The etching reaction was stopped by dilution of the acid solution water is.

The resulting view of the PSi particles was analyzed using Scanning Electron Microscope (SEM), BET and bjh's methods /19/.

2 and 3 - SEM of microsemi particles of porous silicon (PSi)set forth in this example method.

Research BET and bjh's methods were made using a micrometer device /18/ and the procedure described above. Point of adsorption and desorption were taken from 0.01 to 0.99 relative units.

The surface of the particles, based on the BET analysis surface area with 5 points, was in the range from 1,8559 to 0,0124 m2/g with correlation coefficient 0,999926. Accordingly, the surface area of the particles was increased approximately 11 times using the above described method of etching. The average pore diameter was about 4,77075 nm and was determined on the basis of the analysis of the full volume of the pore space, BET specific technology.

The results of adsorption bjh's analysis then listed in Table 1.

Table 1
The diameter of pores

Range (nm)
The average Diameter(nm)Given Pore Volume (cm3/g)The integral Pore Volume (cm3/gGiven the Surface of Pores (cm2g)The integral health of the Surface of Pores (cm2/g)
210,62-295,20238,980,0015510,0015590,0260,026
103,15-210,82123,500,0000710,0016310,0020,028
81,15-103,1589,500,0000270,0016570,0010,030
40,42-81,1548,220,0000770,0017340,006being 0.036
27.18 per-40,4231,200,0000490,0017840,0060,042
20,35-27.18 per22,760,0000310,0018140,0050,048
16,32-20,3517,870,0000410,0018550,0090,057
of 13.58-6,3214,690,0000260,0018810,0070,064
11,31-of 13.5812,230,0000360,0019170,0120,076
10,55-11,3110,900,0000130,0019300,0050,080
8,09 -10,558,970,0000790,0020090,0350,116
8,65-8,097,22,000067 0,0020760,0370,153
5,64-6,656,050,0001560,0022330,1030,256
4,85-5,645,180,0001650,002397to 0.1270,383
4,23-4,854,490,0002240,0026220,2000,583
3,72-4,233,940,0002290,0028510,2320,816
3,29-3,723,480,0002210,0030720,2551,070
2,93-3,29is 3.080,0001930,0032640,2501,320
2,61-2,932,740,0001580,0034220,2301,550
2,32-2,612,440,0001280,0035500,2101,760
2.06 to 2,322,170,0001160,0036660,2141,974
1,82 e 2.061,920,0000950,0037620,1982,172
1,72-1,821,770,0000400,0038020,0922,264

The results of BJ analysis of pore distribution desorption are summarized in Table 2.

Table 2
The pore diameter Range (nm)The average Diameter(nm)Given Pore Volume (cm3/g)The integral Pore Volume (cm3/g)Given the Surface of Pores (cm2/g)The integral Surface of Pores (cm2/g)
278,56-294,91286,270,0011550,0011550,0160,016
87,25-278,56103,770,0005040,0018590,019being 0.036
62,76-87,2571,020,0000400,0017000,0020,038
35,37-62,7641,800,0000650,0017650,0060,044
24,52-35,3727,970,0000500,0018150,0070,051
20,20-24,5221,930,0000330,0018490,0060,057
16,09-20,2017,660,0000380,0018870,009of 0.066
12,69-16,0913,980,0000410,0019280,0120,078
10,89-12,69 11,650,0000260,0019540,0090,087
to 9.91-10,8910,350,0000240,0019780,0090,096
7,81-to 9.918,600,0000590,0020370,0270,123
6,40-7,816,950,0000750,0021120,0430,166
are 5.36-6,405,760,0000960,0022080,0670,233
4,57 and 5.364,900,0001240,0023320,1010,334
3,95-4,574,210,0001840,0025150,1740,508
3,44-3,953,660,0002220,0027380,2430,752
3,01-3,443,200,0002700,003008of 0.3371,089
2,65-3,012,810,0002620,0032700,3731,462
2,29-2,652,440,0001720,0034420,2821,744
2,00-to 2.292,120,0001200,0035620,2261,70
1,74-2,001,850,0001010,0036630,2192,189

From the above results it is clear that there are no clogged or other restraint then analyzed sample. Method bjh's showed that the integral adsorption volume of the pore space of the pores having a pore diameter between 1.7 and 300 nm was 0,003802 cm3/g, while the cumulative volume of pore space desorption was 0,003663 cm3/g. Bjh's integral adsorption surface area of pores having a pore diameter between 1.7 and 300 nm, was 2,2636 m2/g, while the bjh's integral surface desorption was 2.1889 m2/g.

The technique based on the determination of the full volume of the pore space, also defined by a bjh's method gave values of average diameters of the pores of the adsorption and desorption 6,71859 nm and 6,69407 nm, respectively.

Differences in surface area and the values of the pore diameter of between methods of measurement BET and bjh's show that the pores are of a cluster of cylinders. However, SEM of microsemi refute this, they show that the porous structure of the particles cannot be characterized as a cluster of cylinders. Accordingly, the analysis of the surface area BET probably provides a more accurate value of the surface area.

Example 2 - Production of PSi cha is TIC

PSi particles were prepared as in Example 1, except. that silicon particles were suspended in the acid solution for 30 seconds. Received PSi particles was analyzed by BET analysis.

BET analysis was performed by the instrument /18/using the procedure described above. The amount of absorbing particles were taken from 0.05 to 0.3 relative units.

The BET surface area of the particles, obtained on the basis of the analysis BET surface area of from 5 points, ranged from 0,6858 to 0.0039 m2/g with correlation coefficient 0,999934. Accordingly, the surface area of the particles was increased approximately 4 times using the above technique of etching.

Figure 4 and 5 shows SEM of microsemi PSi particles produced by the method of this example.

Example 3 Oxidation of PSi particles peroxide

PSi particles produced in Example 1 were subjected to chemical oxidation using hydrogen peroxide. PSi particles was maintained at room temperature for 1 hour in 30% aqueous peroxide solution.

Example 4 Oxidation of PSi particles using dimethyl sulfoxide (DMSO).

PSi particles produced in Example 1 were subjected to chemical oxidation, using only the sulfoxide (DMSO) and DMSO solution containing 500 mg/ml 2,6-di-tert-butyl-4-METHYLPHENOL (EIT), acceptor of free radicals. PSi particles vydergivanija room temperature for 1 hour in DMSO and 2 hours in a solution of DMSO/BHT.

Example 5 Oxidation of PSi particles using iodine crystals

PSi particles produced in Example 1 were subjected to chemical oxidation using iodine crystals. PSi particles were aged in the presence of iodine crystals: (1) in vacuum and (2) in the air.

In vacuum, the particles were kept in a thermos containing iodine crystals for 2 hours at room temperature. Then the particles were placed in an air environment at room temperature.

In the air of particles contained in a sealed tube flask containing iodine crystals for 2 hours at room temperature. Then the particles were placed in an air environment at room temperature.

Example 6 - Add the main linker

The oxidized particles prepared in Example 3 were immersed in 10% (by volume) solution of 3-glycidoxypropyltrimethoxysilane, the main linker in water for 4 hours at 75°With, then quickly was subjected to annealing at 110°to form a covalent bond between the 3-glycidoxypropyltrimethoxysilane and hydroxyl groups on the surface of PSi particles.

Example 7 - Add main and secondary (Sulfo-SMCC) linker

The oxidized particles prepared in Example 3 were immersed in 10% (by volume) solution of aminopropyltriethoxysilane, the main linker in water for 4á at 75° With, and then quickly was subjected to annealing at 110°to form a covalent bond between aminopropyltriethoxysilane and hydroxyl groups on the surface of PSi.

The particles were immersed in a solution of sulfosuccinimidyl 4-(N-maleic aminomethyl)-cyclohexane-1-carboxylate (sulfo-SMCC) secondary linker with heterobifunctional communication. The particles were stored in a solution containing 10 mg of Sulfo-SMCC in ml of water for 1 hour at room temperature to form a covalent bond between the main and secondary linkers. After soaking in the Sulfo-SMCC solution particles were washed in water.

Example 8 - Adding the main and secondary (glutaraldehyde) linker

The oxidized particles prepared in Example 3 were immersed in 10% (by volume) solution of aminopropyltriethoxysilane, the main linker in water for 4 hours at 75°With, then quickly was subjected to annealing at 110°to form a covalent bond between aminopropyltriethoxysilane and hydroxyl groups on the surface of PSi particles.

Then the particles were immersed in a solution of glutaraldehyde (2.5% in phosphate buffer), homobifunctional secondary link, the linker, for 1 hour at room temperature to form a covalent bond between the linker and the secondary link Builder. After you remove the key in the solution of glutaraldehyde particles were washed in water.

Example 9 - Adding the main and secondary (BS3) linker

The oxidized particles prepared in Example 3 were immersed in 10% (by volume) solution of aminopropyltriethoxysilane, the main linker in water for 4 hours at 75°With, then quickly was subjected to annealing at 110°to form a covalent bond between aminopropyltriethoxysilane and hydroxyl groups on the surface of PSi particles.

Then for education homobifunctional communication linker particles were immersed in a solution (sulfosuccinimidyl)suberate (BS3at a concentration of 5 mg per 1 ml of water for 1 hour at room temperature to form covalent bonds between the main and secondary linker. After soaking in BS3the solution particles were washed in water.

Example 10 - Add Item Recognition in the form of Antibodies

Purified body of immunoglobulin G(IgG)obtained from /20/, were attached to the main linker (3-glycidoxypropyltrimethoxysilane) of the particles produced in Example 6 exposure IgG in phosphate buffered saline solution, at 37°C for 90 minutes.

For visualization and quantification fixing antibodies specific fragment of the antibody immunoglobulin IgG F (AB’)2associated with SS3 fluorescent marker (krasnayapresnya, generated from /20/, was introduced into contact with the particles having an attached antibody IgG. Fluorescently-labeled fragment of the antibody immunoglobulin IgG F (ab’)2in phosphate buffered saline solution was maintained for 30 minutes at room temperature in the presence of particles with bound IgG. The complex structure of the agreed fixed body of IgG and fragments of antibodies IgG F (ab’)2was investigated using the fluorescent epichlorhydrine microscopy and quantitatively measured by fluorometry. Figure 6 shows a fluorescent micrograph obtained by the microscope Nikon Diaphot 300 at 200-fold magnification. The picture shows the structure of the interaction of antigens PSi particles, modified IgG antibodies.

Example 11 - Add Item Recognition in the form of Antibodies

Purified body of immunoglobulin G (IgG)obtained from /20/partially processed three-(2-carboxyethyl) hydrochloride hydrogen phosphide (TSER)at room temperature for 25 minutes, was subjected to sulfhydryl groups of the antibody. Specified IgG was attached via a sulfhydryl reactive group to the secondary communication linker (sulfo-SMCC) particles produced in Example 7 exposure specified IgG in phosphate buffered saline solution at 37°C for 0 minutes.

For visualization and quantification fixing antibodies specific fragment of the antibody immunoglobulin IgG F (ab’)2associated with SS3 fluorescent marker, was injected into contact with the particles having an attached antibody IgG. Fluorescently-labeled fragment of the antibody immunoglobulin IgG F (ab’)2in phosphate buffered saline solution was maintained for 30 minutes at room temperature in the presence of particles with bound IgG. The complex structure of the agreed fixed body of IgG and fragments of antibodies IgG F (ab’)2was investigated using the fluorescent epichlorhydrine microscopy and quantitatively measured by fluorometry. She showed good coverage and distribution of item detection antibodies on the PSi particles.

Example 12 - Add Item Recognition in the form of Antibodies

Purified body of immunoglobulin G (IgG)obtained from /20/, were attached to the secondary communication linker (aldehyde of glutarate) particles produced in Example 8 exposure specified IgG in phosphate buffered saline solution at 37°C for 90 minutes.

For visualization and quantification fixing antibodies specific fragment of the antibody immunoglobulin IgG F (AB’)2associated with SS3 fluoresc ntim marker, was introduced into contact with the particles having an attached antibody IgG. Fluorescently-labeled fragment of the antibody immunoglobulin IgG F (ab’)2in phosphate buffered saline solution was maintained for 30 minutes at room temperature in the presence of particles with IgG attached.

Comparable results were obtained for the oxidized particles produced in Examples 4 and 5 and provided with main and secondary linker in Example 8.

Example 13 - Adding Enzyme Item Recognition

The enzyme acetylcholinesterase /20/ was attached via a secondary communication aldehyde of glutarate linker particles produced in Example 8 exposure of the enzyme in phosphate buffered saline solution at room temperature for 90 minutes.

7 graphically compares the control enzyme, obtained by the hydrolysis of acetylcholine by the enzyme caused by enzymatic modification of PSi particles.

Example 14 Treatment of protein A, adding the body of IgG antibodies

Particles produced in Example 8, were treated with protein by exposure of the particles in the solution of protein in phosphate buffered saline solution at 37°C for 3 hours. After cooling, the particles were washed with phosphate solution with a buffer additive.

Purified body, antic the La immunoglobulin G (IgG), generated from /20/, were attached to the protein A. the Treated particles were stored in phosphate buffered saline IgG at 37°C for 90 minutes.

For visualization and quantification fixing antibodies specific fragment of the antibody immunoglobulin IgG F (ab’)2associated with SS3 fluorescent marker, was injected into contact with the particles having an attached antibody IgG. Fluorescently-labeled fragment of the antibody immunoglobulin IgG F (ab’)2in phosphate buffered saline solution was maintained for 30 minutes at room temperature in the presence of particles with IgG attached.

Fluorescent epichlorhydrine microscopy and fluorometry showed good coverage and distribution of item detection antibodies on the PSi particles.

Also used the analysis associated with fluorescent markers to determine minimized if the protein And non-specific fixation of biomolecules to processed protein And the surface of the PSi particles, compared to the immunoglobulin used in the examples described above. And fluorescent epichlorhydrine microscopy and fluorometry showed no significant non-specific fixation of biomolecules other than immunoglobulin to intact Fc domains.

Example 15 - Education is denied protein And, adding Collagen IV

Particles produced in Example 8, were treated with protein, by keeping the particles in a solution of protein in phosphate buffered saline solution at 37°C for 3 hours. After cooling, the particles were washed with a phosphoric acid solution with a buffer additive.

Purified antibodies collagen IV derived from /20/, were attached to the protein A. the Treated particles were stored in phosphate buffered saline IgG at 37°C for 90 minutes.

For visualization and quantification fixing antibodies specific fragment of the antibody immunoglobulin IgG F (AB’)2associated with SS3 fluorescent marker, was injected into contact with the particles having an attached antibody IgG. Fluorescently-labeled fragment of the antibody immunoglobulin IgG F (ab’)2in phosphate buffered saline solution was maintained for 30 minutes at room temperature in the presence of particles with IgG attached.

Fluorescent epichlorhydrine microscopy and fluorometry showed good coverage and distribution of item detection antibodies on the PSi particles.

Also used the analysis associated with fluorescent markers to determine minimized if the protein And nonspecific fixation biome is the molecules to processed protein And the surface of the PSi particles compared with immunoglobulin, used in the examples described above. And fluorescent epichlorhydrine microscopy and fluorometry showed no significant non-specific fixation of biomolecules other than immunoglobulin to intact FC domains.

Example 16 Treatment of the blocking solution

To minimize nonspecific fixation using the blocking solution was tested to contain particles produced in Examples 6, 8 and 9. The particles were treated with a buffer solution aminouksusnoy acid (or 50 mm, or 200 mm, or at a pH of 8.6, or pH 10) for 30 and 60 minutes at room temperature. After cooling of the particles in the buffer solution aminouksusnoy acid body IgG (used here as a test material having amine reactive groups)associated with SS3 fluorescent marker, were decorated with particles at 37°C for 90 minutes. After cooling, the particles were washed in phosphate buffered saline solution. To determine the impact of blocking buffer aminouksusnoy acid, i.e. the ability of a buffer to slow the consolidation of the protein on the linker was made by evaluation of fluorescently-labeled IgG deposited on the particles. And fluorescent epichlorhydrine microscopy and fluorometry showed a significant, >70%, the fixing antibodies in the treated PSi particles. Similar results would be and obtained for particles produced in Examples 6, 8 and 9.

On Fig presents a fluorescent micrograph of 200-fold magnification, which shows the fixing antibodies in PSi particles that are not processed in aminouksusnoy acid. Figure 9 presents a fluorescent micrograph of 200-fold magnification, which demonstrated a decrease in the number of attached antibodies in PSi particles treated with buffer aminouksusnoy acid.

Practically, the recognition elements were attached to the PSc particles before treatment with blocking solution. The blocking solution minimizes the consolidation of non-specific compounds and analyzed subject directly to the surface of the PSc.

To verify that the buffer aminouksusnoy acid has no adverse effect on the ability of IgG to interact with antigen-specific antibody fragment of the antibody immunoglobulin IgG F (ab’)2associated with fluorescent ether of sortanova acid, fluorescent marker was added to the IgG for 30 accurate exposures at room temperature. Binding activity of IgG, following exposure in the buffer aminouksusnoy acid, was evaluated by fluorescent epichlorhydrine microscopy and fluorometry. No harmful effects of buffer aminouksusnoy acid on a consistent consistent binding activity of IgG was observed.

Figure 10 p is edstaven fluorescent micrograph of 200-fold magnification, which demonstrates that the buffer aminouksusnoy acid does not adversely affect the activity of antibodies in Fig.9.

Example 17 - a study of the Photoluminescence

The device shown in figure 1, was used to analyze the photoluminescence of many measurements made in the above Examples. Light having a wavelength of 488 nm, was focused on the sample. The spectral sensitivity of the photodiode was in the range of from about 420 nm to 1100 nm.

Photoluminescence PSi particles created in Example 1, was analyzed, and the PL intensity is graphically represented as a function of photon energy and wavelength figure 11. PL peak intensity 625 relative units, which corresponded approximately to the level of 1.94 eV.

Photoluminescence PSi particles created in Example 8, was analyzed. PL intensity graphically represented as a function of photon energy and wavelength on Fig. PL peak intensity was approximately 1000 relative units, which corresponded approximately to the level of 2.24 eV.

Photoluminescence PSi particles with item recognition in the form of antibodies produced in Example 8, was also analyzed. The analysis of this radiation is determined that the PL intensity is graphically represented as a function of photon energy and wavelength figure 3. PL peak intensity was approximately 1900 relative units, which corresponded approximately to the level of 2.27 eV.

The results of photoluminescence show a significant, explicit and reproducible PL modulation. The expected fluorescence signal of lower energy in the orange-red region of the visible part of the spectrum emitted by the PSi particles of Example 1, shows the increase of energy in the yellow region of the spectrum, adding the element of recognition and explicit more than the high-energy signal in the green region of the spectrum produced by adding the analyzed subject to item recognition. Emitted by the photoluminescent radiation measured used by the device (Figure 1), had an intensity sufficient to distinguish by eye, there was a strong visible luminescence.

We have described above the preferential embodiments of the invention. It is clear that the preceding examples are only part of the possible implementations, but other implementations can be used without going beyond the scope of the proposed claims.

INDUSTRIAL APPLICABILITY

Patterns PSc/RE can be used both in isolation and in combination with a wide variety of devices that support a variety of applications, including, without limitation, biotechnology type is biosensory for medicine, protection and control environment for industrial applications. For the purposes of protection (for example, for chemical and biological detection/identification of factors harmful effects), genetic applications, diagnostics and other molecular/cell biological questions such as selection/sorting cells, microstructural analysis of cells, etc.

Sources of information

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2. Schmuki et al Appl Phys Lett 72:1039; 1998.

3. Patent CNos. 5,338,415 (Sailor et al).

4. Patent Ns. 5,453,624 (Sailor et al).

5. Science 278:840; 31 October 1997.

6. JAm Chem Soc 120:12108-12116; 1998.

7. Bressers et al J Electro-analytical Chemistry, 406:131; 1996.

8. Santa Clara, California, USA.

9. International publication of PCT application W098/25090 (Ishikawa. June 11, 1998).

10. Canham Appl Phys Lett 57:10:1046-1048; 1990).

11. Sailor et al Adv Mater 9:10:783-793; 1997.

12. Kurmaev et al J. of Physics-Condensed Mater 9:2671; 1997.

13. Yamada et al Japanese J Appl. Physics Part I - Regular Papers Short Note 35:1361:1996.

14. Schmuki et al Phys Rev Lett 80:18:4060-4063; 1998.

15. Tsai et al Appl Phys Lett 60:170; 1992.

16. Fauchet J Luminescence 70, 294; 1996.

17. Liu et al, Solid State Communications 106:211; 1998.

18. ASAP 2000TMinstrument (Norcross, Georgia, USA).

19. Bjh's (Barett-Joyner-Halenda.

20. Sigma-Aldrich Canada Ltd. (Oakville, Ontario, Canada).

1. The composition of the modified semiconductor including at least one semiconductor material having a porous texture, and at least one element recognition, modifying the specified semiconductor material, and ensuring the maintenance, at least one first luminescent reaction in the range of about 200 to 800 nm when covering songs by electromagnetic radiation, at least one wavelength in the range of about 100 to 1000 nm.

2. The composition according to claim 1, characterized in that the element recognition selected from the group consisting of biomolecular components, organic components, inorganic components and their combinations.

3. The composition according to claim 2, wherein the biomolecular components selected from the group consisting of natural or synthetic proteins, nucleic acids, oligonucleotides, lectins, carbohydrates, glycoproteins, lipids and combinations thereof.

4. The composition according to claim 1, characterized in that it contains at least one item analysis, interacting with the composition of semiconductors.

5. The composition according to claim 4, characterized in that the object of analysis selected from the group consisting of inorganic, organic and biomolecular structures.

6. The composition according to claim 4, characterized in that at least part of the specified subject of the analysis interacts with the specified composition modified semiconductors.

7. The composition according to claim 4, characterized in that at least part of the specified subject of the analysis interacts with the specified poluprovodnikov material.

8. The composition according to claim 4 characterized in that that, at least some of the listed item analysis interacts with the specified element of recognition.

9. The composition according to claim 4, characterized in that when the lighting of the specified composition of semiconductors are the subject of analysis of at least one wavelength of electromagnetic radiation in the range of about 100 to 1000 nm, the semiconductor composition/subject analysis produces at least one second luminescent reaction in the range of about 200 to 800 nm.

10. The composition according to claim 9, characterized in that the at least one second luminescent reaction is modulated, at least in intensity or wavelength compared with the specified at least one first luminescent reaction.

11. The composition according to claim 1, characterized in that the largest average size of the specified semiconductor material is in the range of about 100 nm - 1 mm

12. The composition according to claim 1, characterized in that the element recognition covalently associated with the specified semiconductor material.

13. The composition according to claim 1, characterized in that it contains at least one main linker between the specified element recognition and specified the semiconductor material.

14. The composition according to item 13, characterized in that it contains at least one secondary communication comp is NOVICA between the specified major linker and the specified item of recognition.

15. The composition according to item 13, wherein the composition contains processing linker.

16. The composition according to 14, characterized in that it contains the composition processing of the linker.

17. The composition according to item 15, wherein the composition processing linker selected from the group consisting of immunoglobulin proteins, bilinovich reactive means of blocking solutions and their combinations.

18. The composition according to item 16, characterized in that the composition processing linker selected from the group consisting of immunoglobulin proteins, bilinovich reactive means of blocking solutions and their combinations.

19. The composition according to claim 1, characterized in that the semiconductor material is selected from the group consisting of silicon, silicon carbide, silicon dioxide, germanium, gallium, gallium arsenide, silicon phosphide, cadmium, selenium, copper oxide, and combinations thereof.

20. The composition according to claim 1, characterized in that said semiconductor material contains an impurity.

21. The composition according to claim 20, wherein the impurity is selected from the group comprising erbium, boron, Fosforit, copper, phosphors of the series of lanthanides, and combinations thereof.

22. The composition according to claim 1, wherein the semiconductor material includes additional material active zone, as the basis for a given semiconductor material.

23. Comp is the position of item 22, characterized in that the material of the active zone is selected from the group consisting of glass, plastics, ceramics, zeolites, metals and combinations thereof.

24. The composition according to claim 4, characterized in that the object of analysis is in the body.

25. The composition according to claim 4, characterized in that the object of analysis is obtained from an organism.

26. The method of measurement of fluorescent reaction in the analysis of substances, including coverage of electromagnetic radiation, at least one wavelength in the range of about 100 to 1000 nm compositions modified semiconductor consisting of a semiconductor material having a porous texture, and at least one element recognition, modifying the specified semiconductor material, and providing at least one first luminescent reaction in the range of 200 to 800 nm at the specified coverage, and measuring at least the intensity or wavelength of the specified at least first fluorescent response.

27. The method according to p, characterized in that the contacting of the specified composition of the modified semiconductor with the subject of the analysis with the formation of semiconductor and analytical compositions, the element of recognition is a tool with the subject of the analysis specified illumination by electromagnetic radiation, the edge is her least one wavelength in the range of about 100 to 1000 nm is at least the specified composition of semiconductors and subject analysis with obtaining at least one second luminescent reaction measurement, at least, intensity, or wavelength, of the specified at least one second luminescent reaction, comparison, at least, intensity, or wavelength mentioned at least first and second luminescent reactions to determine the modulation between at least one of the first and second luminescent reactions and based on a specified comparison is the presence of the specified subject of analysis.

28. The method according to item 27, wherein the subject of the analysis is in the body.

29. The method according to item 27, wherein the subject of analysis is obtained from an organism.

30. The method according to item 27, wherein the analysis is selected from the group consisting of inorganic, organic and biomolecular structures.



 

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FIELD: devices built around diodes emitting blue and/or ultraviolet light.

SUBSTANCE: proposed light source emitting light in ultraviolet or blue light region (from 370 to 490 nm) and capable of producing high-efficiency white light affording control of luminance temperature within comprehensive range has light-emitting component that emits light in first spectral region and phosphor of group of optosilicate alkali-earth metals and that absorbs part of source light and emits light in other spectral region. Novelty is that phosphor used for the purpose is, essentially, europium activated bivalent optosilicate of alkali-earth metal of following composition: (2-x-y)SrO · x(Bau, Cav)O · (1-a-b-c-d)SiO2 ·aP2O5bAl2O3cB2O3dGeO2 : yEu2+ and/or (2-x-y)BaO · x(Sru, Cav)O · (1-a-b-c-d)SiO2 ·aP2O5bAl2O3cB2O3dGeO2 : yEu2+.

EFFECT: enhanced efficiency, enlarged luminance temperature control range.

14 cl, 10 dwg

FIELD: semiconductor emitting devices.

SUBSTANCE: proposed light-emitting diode based on nitride compounds of group III metals, that is aluminum, gallium, and indium (AIIIN), includes p-n junction epitaxial structure disposed on insulating substrate and incorporating n and p layers based on solid solutions of group III nitrides AlxInyGa1 - (x + y)N, (0 ≤ x ≤ 1, 0 ≤ y ≤ 1), as well as metal contact pads for n and p layers disposed on side of epitaxial layers, respectively, at level of lower epitaxial n layer and at level of upper epitaxial p layer. Projections of light-emitting diode on horizontal sectional plane, areas occupied by metal contact pad for n layer, and areas occupied by metal contact pad for p layer are disposed on sectional plane of light-emitting diode in alternating regions. Metal contact pad for n layer has portions in the form of separate fragments disposed in depressions etched in epitaxial structure down to n layer; areas occupied by mentioned fragments in projection of light-emitting diode onto horizontal sectional plane are surrounded on all sides with area occupied by metal contact pad for p layer; fragments of metal contact pad for n layer are connected by means of metal buses running over metal contact pad insulating material layer applied to portions of this contact pad over which metal buses are running.

EFFECT: enhanced output optical power and efficiency of light-emitting diode.

3 cl, 3 dwg

The invention relates to semiconductor devices intended to emit light, namely light-emitting diodes containing a p-n junction as the primary radiation source

The invention relates to the field of semiconductor emitting devices

Led lamp // 2231170
The invention relates to lighting devices and can be used, in particular, when designing the lighting lamps in medical technology

The light source // 2220478

The invention relates to optoelectronics, namely the structures of the semiconductor radiation sources white

The invention relates to solid-state electronics, in particular, to designs of powerful light sources based on semiconductor emitting crystals

The light source // 2210143
The invention relates to light sources and can be used in lighting equipment, systems for regulating the movement of vehicles, in particular water transport

FIELD: medicine.

SUBSTANCE: method involves measuring luminescence parameters in to areas on biological object (control area and case one). Each area is exposed to optical radiation action sequentially in spectrum segments corresponding to various tissular fluorophors luminescence excitation. Radiation components are selected from luminescence radiation caused by optical treatment applied to the areas on biological object in each of mentioned spectrum segments. Their intensity is measured synchronously with appropriate pulsation wave phase in corresponding biological object area. Device is usable for excluding artifact and general somatic state influence upon luminescence parameters measurement results.

EFFECT: high accuracy in estimating fluorescence properties of aerobic and anaerobic bacteria.

27 dwg, 3 tbl

FIELD: measurement technology.

SUBSTANCE: glowing of tested object is excited and pulses are amplified, formed, registered and compared with test-object. Tested object is subject preliminary to freezing, then the object is placed inside container made of low heat conductivity material. Container is placed into light-tight chamber provided with shutter. After glowing is established, quantum light flux radiated by tested object is passed through optical mirrors located at input of photomultiplier tube. Glowing is initiated by flash light due to contact tool which makes photo flash connection circuit synchronously with operation of shutter.

EFFECT: improved precision of measurements; improved precision.

1 dwg

FIELD: analysis of water and organic solutions.

SUBSTANCE: sensor has multi-channel structure in form of length 1 of poly-capillary pipe with through capillary, forming micro-channels, which are filled with two layers of non-mixing substances. One layer 4 is formed by water or water solution and other 3 - by organic substance. In first of said layers into micro channels micro-granules 5 of absorbent are placed.

EFFECT: higher efficiency, lower costs.

27 cl, 14 dwg

FIELD: analytical methods.

SUBSTANCE: method is based on specific physicochemical feature of combination of impurity traces, which determine composition of product, said feature being capacity of absorbing and reemitting optical emission (luminescence). According to invention, one accumulates files of spectral-luminescent characteristics of product being identified and reference product. Identification procedures allow one to follow minor deviations of characteristics in liquid composition.

EFFECT: increased identification efficiency.

5 dwg, 4 tbl

FIELD: medicine.

SUBSTANCE: method involves taking bone tissue fragment sample in area under examination, measuring relative laser luminescence level. The obtained values are compared to normal bone tissue characteristics. Quantitative reduction of mineral composition being found relative to reference value in normal state is diagnosed by interpreting spectral characteristics in diagnostic bandwidth of 350-550 nm.

EFFECT: high accuracy of diagnosis.

2 dwg

The invention relates to the field of optical methods

The invention relates to measuring equipment

The invention relates to analytical chemistry, namely the manufacture of fluorescent sensor for determining the oxygen content in gases and liquids over the entire range of oxygen concentrations and can be used in analytical chemistry, chemical and food industry, medicine, biotechnology, environmental monitoring, environmental

The invention relates to optical methods of measurement of physico-chemical parameters

FIELD: analytical chemistry of elements.

SUBSTANCE: the invention is pertaining to the field of analytical chemistry of elements, in particular, to methods of detection of silver availability, that may be used at detection of silver availability in the natural waters and technological solutions. The method of detection of silver availability provides for preparation of a silver solution (1), its transformation into a complex compound and measurement of the luminescent emission intensity. Silver is separated from solutions using a silica gel chemically modified with the help of mercaptogroups and intensity of the silver complex compound luminescent emission (1) is measured with the help of mercaptogroups on the surface of silica gel at 77К at ultraviolet irradiation light. The technical result is simplification of the technological process, decrease of a relative limit of detection, extension of the range of defined concentrations.

EFFECT: the invention ensures simplification of the technological process, decrease of a relative limit of detection, extension of the range of defined concentrations.

3 ex

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