The method of preparation of bone samples for morphological diagnosis

(57) Abstract:

The invention relates to medicine, namely morphology, can be used for morphological diagnosis of pathology of the bone tissue. The essence of the method: after fixation of bone samples in formalin solution are declinatio within 7-8 days in a 5% solution of nitric acid on the basis of a 5% formalin solution, after which bone samples for 2 h, washed under running tap water and 12 h immersed in a 5% solution of sulfuric acid lithium, and within 20-30 min, again washed with running water and subjected to dehydration in alcohols of increasing concentration. The proposed method reduces the processing time of bone samples obtained blocks are more plasticity, which allows to obtain high-quality histological sections, which are characterized by the preservation of all structures of the bone tissue.

The invention relates to medicine, namely morphology, can be used for morphological diagnosis of pathology of the bone tissue.

There are two main groups of methods decalcomanie - acid-free and acid[4, 5, 7, 8, 9]. Acid method decalcomania [2, 3] is to prepare scow 15-20% wage formalin solution not more than 48 hours; 2) washing the bone samples running water for 24 h; 3) in their dehydratation in solutions of alcohols 50%, 70%, 96%, 70%, 50%, during the day in a solution of alcohol each concentration.

Decalcomania:

1) observing the rules of decalcomania, bone samples are placed in a 5-7,5% solution of nitric acid on the basis of distilled water for 10-12 days; 2) then the bone sample is placed in a 5% solution of sulfuric acid sodium or lithium for 24 h to eliminate swelling; 3) bone samples washed in running tap water for 1-2 days; 4) perform fill in celloidin or paraffin or cut on a freezing microtome.

The disadvantages of this method decalcomania are: the length of time decalcomanie (10-12 days). the duration of time washing the bone samples in flowing water (2-3 days) that contribute to the saturation of their liquid, reduces quality of coating sections and clarity manifestations structures of the bone tissue.

The objective of the proposed invention is the reduction of terms of decalcomania, increasing the plasticity of bone samples and the preservation of bone structure.

The task carried out due to the fact that decalcomania to the donkey what their for 2 h, washed under running tap water and 12 h immersed in a 5% solution of sulfuric acid lithium then for 20-30 min, again washed with running water and subjected to dehydration in alcohols of increasing concentration.

The method is as follows.

Bone samples of size 1,0×1.0 cm (cadaveric material) is fixed at 7-8% wage formalin solution for 2 days. They are then washed under running tap water for 20-30 min and immersed in a 5% solution of nitric acid on the basis of a 5% formalin solution [1, 6].

The acid solution on the basis of formalin changed daily. Decalcomania lasts 7-8 days, to determine its end use puncturing samples preprofile needle, a trial cut and clenching. Then bone samples should be rinsed with running tap water for 2 h and immersed for 12 h in a 5% solution of sulfuric acid lithium to remove their swelling.

Next, after washing with running water for 20-30 min, bone samples are subjected to dehydration in alcohols of increasing concentration(50%, 70%, 90%, 96%, absolute alcohol, the alcohol - ether mixture). Each solution sample was incubated for days. Then they should fill in celloidin.

To achieve full decalcomanie allows the solution of nitric acid, prigotovlenn is the shaft of the samples contribute to the reduction of time spent in the solution of sulfuric acid lithium their dehydration just before pouring and pouring in celloidin, which in its properties (plasticity) is close to decalcified bone samples. To assess the completeness of decalcomania and plasticity of bone samples is possible only in the preparation of the slices. There is no crunch and get a full, smooth cuts. Preservation of bone structure can only be assessed after staining sections (Smarly) and histomorphometry. In the studied drugs are defined layers of the bone, otuonye structure of the cell.

The proposed method allows to reduce the duration of decalcomanie 5-7 days to achieve full decalcomania, to increase the plasticity of bone samples to preserve the structure of the bone.

References

1. Alencon B. A. Method of accelerated decalcomania in acid-formalisation solution. Archive pathol. 1950, I. XII, n 5, c.93. 2. Volkova O. C., Eletsky Y. K. Fundamentals of histology with histological technique. - M.: Medicine, 1971, S. 271. 3. Elisha C., fundamentals of histological techniques. - M.: Medicine, 1967, S. 209. 4. Klevezal, A. Registering patterns mammals in Zoological research. - M.: Nauka, 1988, S. 285. 5. Lilly R. Histopathological technique and practical histochemistry. - is licencie teeth and fragments of jaws. Bull. East.-Sibirsk. the scientific. center WITH the RAMS. - Irkutsk, 1993, vol.3-4, S. 150-154. 8. Nikonorov, S., Kozlov I. A. Decalcomania bone in increasing concentrations of salt trylon B. Archive Anat., pistol. and embryol. 1986, so XCL, N. 1, S. 78-80. 9. D. Sarkisov, S., Perov, Y. L. Microscopic techniques. A guide for physicians and laboratory technicians. - M.: Medicine.

The method of preparation of bone samples for morphological diagnosis, including the fixation of bone samples in formalin solution, washing with water, declinatio 5% solution of nitric acid, maintaining a 5% solution of sulfuric acid with sodium subsequent washing with water, characterized in that declinatio hold for 7-8 days in a 5% solution of nitric acid on the basis of a 5% solution of formalin, and then for 2 h, the samples should be rinsed with running tap water and 12 h immersed in a 5% solution of sulfuric acid lithium then for 20-30 min, again washed with running water and subjected to dehydration in alcohols of increasing concentration.