Apparatus for fluorescent spectroscopy of biological tissue

FIELD: physics.

SUBSTANCE: apparatus comprises a fluorescent-reflective spectrometer, which includes an illumination system and spectrometric system connected to a Y-shaped fibre-optic probe. The apparatus is further provided with two channels, one of which intended for supplying liquid to the investigated organ to wash off blood and is connected to a pump, and the other channel is intended for sucking the liquid and blood from the investigated organ and is connected to a pump. Both channels and the distal end of the fibre-optic probe are placed in a ferrule to form a fibre-optic probe. The ferrule is in the form of a metal cylinder with a socket at the end which is adjacent to the investigated organ.

EFFECT: high accuracy and consistency of measurement results, enabling investigation of the heart within the body.

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The invention relates to medicine, in particular to surgery, to medical equipment, namely, devices for fluorescence spectroscopy of biological tissue and can be used for intraoperative assessment of functional and metabolic status of biological tissues, including to determine the state of the myocardium and other organs in ischemic damage.

For intraoperative assessment of functional and metabolic status of biological tissues, including to determine the state of the myocardium and other organs in ischemic damage, can be used fluorescence spectroscopy. The method is based on the phenomenon that at the early stage of ischemia, the accumulation of excessive amounts of NADH in breach of its oxidation to NAD+ through the chain of mitochondrial transport of electrons, which have the ability to glow in the visible spectrum when excited by ultraviolet rays, and the intensity of the glow depends on the location in the oxidized or reduced state. Thus, the phenomenon of autofluorescence fabrics can be used for non-invasive registration of ischemic changes in organs and tissues. Important aspects of this technology are low invasiveness and technical simplicity of the methodology of Enki the viability of the tissue.

A device for laser-induced fluorescence spectroscopy (LIFAS) to determine ischemia and hypoxia in biological tissue (US 6697657 Method and devices for laser induced fluorescence attenuation spectroscopy (LIFAS)) containing lighting and spectrometric systems, fiber-optic harness connected to the light and spectrometric systems, probe, which houses the working end of the fiber optic bundle, and a processor for processing signals, lighting and spectrometric systems. The use of the device provides high accuracy results in the experiments performed ex vivo, for example, on a stand-alone perfusion heart of the laboratory animal.

However, in similar experiments in vivo, when the heart is still in the body, carrying out such measurements difficult or impossible. This is due to the fact that the blood in the operative field, inevitably falls into the gap between the probe and the surface of the examined tissue, this leads to a decrease in the signal due to absorption by the hemoglobin of the exciting and fluorescent radiation and, as a consequence, instability results. The same difficulties arise when conducting measurements in a clinical setting.

Also known spectrometer for fluorescently-reflective biomedical research (KangUk, Papayan, C., Berezin Century B. Petrishchev N. N., Galagoudza M M Spectrometer for fluorescently-reflective biomedical research // Optical magazine. 2013. So 80. No. 1. C. 32-38), which is the closest to the claimed invention and is selected as a prototype.

Known spectrometer for fluorescently-reflective biomedical research includes led illuminator generating radiation in the near ultraviolet region with a Central wavelength of 365 nm, Υ-shaped fiber-optic probe, which unites in its distal part of the fiber illumination channel and a detection channel (measurement channel), as well as the spectrometer and the computer, which registers the spectrum in the range of 350-750 nm.

However, the known spectrometer for fluorescently-reflective biomedical research has the same drawbacks as similar, i.e. functional disabilities that do not allow for the in-vivo experiments.

The technical object of the present invention is to expand the functionality of the device for fluorescence spectroscopy of biological tissue, as well as improving the accuracy and stability of measurement results in the surgical field.

To achieve a technical result, the device for fluorescence spectroscopy of biological tissue containing fluorescence-reflective spectrometer, including lighting and spectrometric systems, each of which is connected to the appropriate output Υ-shaped fiber-optic probe made from the measurement and illumination fibers, forming respectively illuminating and receiving channels, and a computer, according to the invention is further provided with two channels, one of which is for the supply of liquid to the monitoring body for flushing blood and connected to the pump, and the other channel designed for aspiration of fluid and blood from the test body, is connected with the pump, both the channel and the distal end of the fiber-optic probe placed in the tip, forming a fiber-optic probe and the handpiece made in the form of a metal cylinder with a bell on the end adjacent to the studied organ with two fittings designed for connection of pump and pump to the respective channels.

To achieve a technical result in the device for fluorescence spectroscopy of biological tissue channel for the fluid and the channel for aspiration made in the form of two coaxial metal cylinders, the axes of which coincide with the axis of the fiber-optic probe, or a channel for the fluid and the channel for aspiration made in the form of two cylinders, the axis of which is parallel to the axis of fiber-optionscom the probe, and the distal end of the fiber optic probe is additionally placed in thin-walled metal tube.

In comparison with the known analogs of the proposed solution allows you to extend the functionality of the device by conducting experiments ex vivo, that is, on a stand-alone perfusion heart, but also in-vivo experiments, when the heart is in the organism. This is made possible by offering the essential features that not only enhance the functionality of the device, eliminating the dependence of the measurement results from the presence of blood in the operative field, but also to improve the accuracy of diagnosis of ischemic status of individual sections of the studied organs during the operation and stability of the measurement results in the surgical field.

Run a piece of metal (e.g. stainless steel) simplify and reduce the time of the sterilization process.

Thus, the present invention provides the achievement of the task.

The proposed solution is a new, not known in practice, the development of fiber-optic spectrometers, and the set of distinctive features is not necessary in the prior art. The invention is industrially applicable because of the simplicity of design of the disorder and popularity of technological processes of manufacture of individual elements of the device. This solution involves the use of modern materials and manufacturing methods, serial production industry.

The invention is illustrated by drawings, where:

in Fig. 1 shows a schematic diagram of a device for fluorescence spectroscopy of biological tissue with channels for the fluid and aspiration performed in a coaxial manner;

in Fig. 2 shows the tip in cross-section a-A;

in Fig. 3 - range of autofluorescence infarction in the state of perfusion;

in Fig. 4 - range of autofluorescence infarction in a state of ischemia;

in Fig. 5 - dependence of the integral signal autofluorescence in the range 440-460 nm from the time elapsed since the beginning of the experiment when the cyclic ischemia/reperfusion.

Device for fluorescence spectroscopy of biological tissue (Fig. 1) includes a light source 1 and the spectrometer 2, forming together a fluorescently-reflective spectrometer, a pump for supplying fluid 3, the pump suction 4, Υ-shaped fiber-optic probe, one end of which is made in the form of a harness formed of the illuminating optical fibers, and is illuminating fiber channel 5 connected to the light source 1, the other end of the fiber optic probe is formed from the measuring optical fiber and is a foster fiber to the cash 6, coupled with a spectrometer 2, and also contains a channel for the fluid on the inspected body 7 and the aspiration channel 8, which may be in the form of two coaxial metal cylinders, the axes of which coincide with the axis of the fiber optic probe or in the form of two cylinders, the axis of which is parallel to the axis of fiber-optic probe. The device also contains the tip 9, the socket 10, adjacent to the studied organ and executed as a single unit with the tip 9, the computer 11. The distal end 12 of the fiber-optic probe is made in the form of the loom with the hexagonal packing of six illuminating optical fibers, the center of which is located a measuring optical fiber, and placed in the tip 9. At the tip 9 are also channel the fluid 7 and the aspiration channel 8, which together with the distal end 12 of the fiber-optic probe, optionally placed in thin-walled metal tube to form a fiber-optic probe. The cannula 13 and 14 (Fig. 2) designed to connect a channel for the fluid 7 and the aspiration channel 8 respectively to the pump 3 and the pump 4 through the nozzles 15 and 16 (Fig. 2), which is provided with a lug 9. The connection of lighting fiber channel 5 and the receiving fibre channel 6, respectively, with the light source 1 and the spectrometer 2 is carried out using connector is, for example SMA 905. As of the luminaire 1 can be used, for example, led illuminator own elaboration on the basis of a powerful led LEDEngin LZ1-00U600. As a spectrometer 2 is used, for example, the spectrometer Avantes-2048-USB2, as the pump 3 used peristaltic pump.

Device for fluorescence spectroscopy of biological tissue works as follows.

Fiber-optic probe is superimposed on the monitoring body of the socket 10, which creates a vacuum in the area of contact with the surface of the examined body. The generated vacuum ensures a reliable contact of the distal end of the fiber-optic probe with the body, and fluid flow for flushing the blood prevents the leaking of blood into the gap between the distal end of the probe and the surface of the examined body. After installation of fiber-optic probe, the monitoring body is simultaneously supplied stimulating fluorescence radiation with a wavelength of 365 nm from a light source 1, the illumination channel 5 fiber-optic probe, and a fluid (water or saline) from the pump 3 through the cannula 13 and channel 7 for flushing blood from the surface of the examined body and is made to aspirate fluid and blood through the channel 8, the cannula 14 with a pump 4. In the tissue of the organ to be investigated is excited fluorescence of various endogenous f is uorophores, including NADH. Fluorescent radiation fabric is perceived by the receiver channel 6, which passes it to the input of the spectrometer 2, which is analog-to-digital conversion received by the receiving channel 6 signal fluorescence excitation, primary processing and transmission to the computer 11 via the USB2 interface.

The use of a device for fluorescence spectroscopy of biological tissue can be illustrated by the example of one of the experiments studies of various animal organs, particularly the heart of the rats spent in the fgbi "Almazov center them. C. A. Almazov" of the Ministry of health of Russia. The experiment was to study the dynamics autofluorescence heart during execution of repeated episodes of brief ischemia and reperfusion, known as preconditioning (adaptive phenomenon, which consists in increasing the resistance of the myocardium to the subsequent duration of ischemia). Experience in-vivo were carried out on isolated perfusion on Langendorff heart rats using the layout of the present invention in the mode of continuous registration of an integrated fluorescence intensity at the maximum emission of NADH at a wavelength of 450±10 nm at excitation 365 nm. Fiber-optic probe at the Desk were in constant contact with a working heart. The result is Alicia in fiber-optic probe channel for the fluid and aspiration channel, continued destruction of blood at the site of contact of the probe with the surface of myocard that provided the measurements are independent from the presence of blood in the operative field. Thus it was possible to register the fluorescence spectra of the myocardium in the state of perfusion (Fig. 3) and ischemia (Fig. 4), which shows an increase in signal intensity of autofluorescence in the field of 440-460 nm ischemia. In addition, failed to register the dependence of the integral signal autofluorescence in the range 440-460 nm from the time elapsed from the beginning of the experiment (Fig. 5). The experiment was conducted three cycles of ischemia 17, after which followed a period of reperfusion 18 (Fig. 5). The experiment lasted for 11 minutes. Ischemia was induced in a time period 2:45-3:45, 5:45-6:45 and 8:45-9:45. In Fig. 5 there is a clear correspondence between periods of ischemia and reperfusion and the level of the signal from the spectrometer.

Thus, the use of the claimed invention allows to extend the functionality of a device for fluorescence spectroscopy of biological tissue, as well as to improve the accuracy and stability of measurement results in the surgical field.

1. Device for fluorescence spectroscopy of biological tissue containing fluorescently-reflective spectrometer, including lighting and spectrometric system, each of Kotor is x connected to the appropriate end of the branched part of the Υ-shaped fiber-optic probe, made from the measurement and illumination fibers, forming respectively illuminating and receiving channels, and a computer, characterized in that it is further provided with two channels, one of which is for the supply of liquid to the monitoring body for flushing blood and connected to the pump, and the other channel designed for aspiration of fluid and blood from the test body, is connected with the pump, both the channel and the distal end of the fiber-optic probe placed in the tip, forming a fiber-optic probe and the handpiece made in the form of a metal cylinder with a bell on the end, adjacent to the studied organ with two fittings designed for connection of pump and pump to the respective channels.

2. Device for fluorescence spectroscopy of biological tissue under item 1, characterized in that the funnel with a tip made as a single unit.

3. Device for fluorescence spectroscopy of biological tissue under item 1, characterized in that the channel for the fluid and the channel for aspiration made in the form of two coaxial metal cylinders, the axes of which coincide with the axis of the fiber optic probe.

4. Device for fluorescence spectroscopy of biological tissue under item 1, characterized in that the channel for the fluid and the channel for ASPI is then made in the form of two cylinders, axis which is parallel to the axis of fiber-optic probe.

5. Device for fluorescence spectroscopy of biological tissue under item 1, characterized in that the distal end of the Υ-shaped fiber-optic probe, made in the form of the loom with the hexagonal packing of six illuminating optical fibers, the center of which is located a measuring optical fiber.

6. Device for fluorescence spectroscopy of biological tissue under item 1, characterized in that the ends of the branched part of the Υ-shaped fiber-optic probe connected respectively with lighting and spectrometric systems through connectors.

7. Device for fluorescence spectroscopy of biological tissue under item 1, characterized in that the distal end of the fiber optic probe is additionally placed in thin-walled metal tube.



 

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