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Morphological methods of research

bioptaty a mucosa of antral department of a stomach fixed in 10 % neutral formalin on Lille, carried out conducting on alcohols of increasing concentration, inkubirovali in Chloroformium, paraffin, filled in in paraffin.

At carrying out of researches used following methods:

• a histological estimation of a mucosa of a stomach

• argirofilnyj method Grimeliusa for the general identification of population of neuroendocrinal cells

• immunohistochemical techniques with use of antibodies

To a proliferative nuclear cellular antigen (PCNA) and р53

Microscopical researches spent in light microscope Nikon Eclipse Е400.

For a histological estimation bioptatov in the thickness 5 microns painted sections a hematoxylin, eosine, alcian dark blue.

The estimation histological structures of a mucosa of a stomach was spent according to «the Modified Sydney system of classification and gradation of a gastritis» (1996) with use vizualnoyoanalogovoj scales. At the tissue specimen description morphological changes were estimated polukolichestvenno

(Weak, moderated, expressed) with the indicating of following signs:

• nejtrofilnaja infiltration

• limfoplazmotsitarnaja infiltration

• an atrophy

• an intestinal metaplasia (with the indicating of its type)

Presence in cells of a mucosa of a stomach of markers of an apoptosis (a protein р53) and proliferations (PSNA) defined on paraffinic sections avidin-biotinovym immunoperoksidaznym a method with use of mouse monoclonal antibodies of firm Novocastra, clone DO-7

In delution 1:40 for revealing р53 and clone РС-10 in delution 1:300 for PSNA. It was spent high-temperature demaskirovka an antigen in the citrate buffer.

Quantitative morphological researches are executed by means of system of the computer analysis of microscopical images Nikon nuclear heating plant-1 version 2.12 with use of the program the VIDEO TEST MORPHOLOGY version 4.0.

The estimation of following structures of a mucosa of a stomach was made:

• cells of inflammatory infiltration (lymphocytes, plasmocytes, neutrophils) with calculation of their total and percent of the area occupied with them in sight at augmentation *200

• total of neuroendocrinal cells, percent of the area occupied with them, size of their optical density at augmentation *400

• proliferative activity of cells on degree immunohistochemical ekspresii PCNA with calculation of percent of the occupied area and optical density at augmentation *400

• an apoptosis on degree immunohistochemical ekspresii р53

With calculation of percent of the occupied area and optical density at augmentation *400

The relative area of an expression will give representation about spatial distribution of studied factors. The optical density (brightness unit) reflects concentration of an investigated biological product in object on brightness of a staining and testifies to activity of the studied phenomenon.

Calculation of corresponding structures spent in 3 fields of vision then the average size was calculated.

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A source: ANTSIFEROV Roman Vladimirovich. CLINICO-MORPHOLOGICAL FEATURES of the CURRENT of the ATROPHIC GASTRITIS At PERSONS of the SENIOR AGE GROUPS. The DISSERTATION on competition of a scientific degree of the candidate of medical sciences. St.-Petersburg - 2007. 2007

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