Method for preparing oocyte paraffin cut film

Abstract

The invention provides a method for preparing a paraffin section of an oocyte. The method of the invention comprises the steps of fixing, dehydrating, transparency processing, paraffin embedding and slicing, etc., the method of the invention leads the oocyte to be transferred in the circumstance of liquid during the processes of fixing, dehydrating, transparency processing, etc. without a direct effect from outer force, thus the original morphosis of the oocyte can be maintained in maximum extent; a colorless transparent vessel is used for being filled with the oocyte, to lead the oocyte to be seen clearly; after the oocyte is transferred to a mould box, a xylene is removed to reveal the oocyte, thus the oocyte can be found more easily; before the paraffin embedding, a plurality of colored granules are scattered around the oocyte, thereby being convenient for the location of the oocyte in the paraffin. The method of the invention can make a paraffin section of a few or even a single oocyte of cursorial bird with the diameter of 100 microns to 600 microns, the structure of the gained section is clear and complete, and the staining effect is good.

Description

technical field [0001] The invention relates to a method for preparing paraffin slices, in particular to a method for preparing paraffin slices of oocytes, and the method of the invention is particularly suitable for the preparation of poultry ovary cell slices. Background technique [0002] Immunohistochemistry, also known as immunocytochemistry, refers to the method of using the principle of immunology to examine the in situ antigen or antibody components on cells and tissues through specific antigen-antibody binding reactions and the visible chromogenic substances on antibody labels. . It can identify and locate various tissue components in cells, such as enzymes, polypeptides, nucleic acids, polysaccharides, hormones, etc. Finally, through specific staining, the exact position in the cells can be observed under an optical microscope or a fluorescent microscope. Cell spectrophotometer, The image analyzer can perform in situ quantitative determination of the test substanc...

AI Technical Summary

Problems solved by technology

With the development of cell biology and the advancement of in vitro culture technology, the growth and development of oocytes are studied more and more through cell culture. For the structure of oocytes, especially the products and regulatory hormones of some gene expression in the process of growth and development The role of the substance-like substances is one of the hotspots of people's research. In the past, the research on the development of oocytes usually used slices of ovarian local tissue to obtain the growth and development information of oocytes. The advantage of sliced ​​local ovarian tissue is that one slice can Contains a lot of oocytes at different developmental stages. The disadvantage is that in addition to the oocytes, there are other components such as loose connective tissue. During the dehydration process of the sample, the properties of various tissues contained in it are not uniform, resulting in shrinkage. The degree is inconsistent, and finally the oocytes are deformed, unable to maintain their original morphological structure; and a large amount of expensive antibodies will be consumed in immunohistochemical detection, which will not only fail to enhance the specific detection effect of oocytes, but will lead to non-oval cells Blast-specific staining occurs, increasing the level of background interference

If you want to study the role of an important regulatory factor in female poultry oocytes, you must kill multiple female birds at different times to take samples by using the method of ovary sectioning. Individual targeting is not strong, the accuracy of detection is not high, and the confidence interval becomes wider

[0004] Now the technology and conditions of in vitro cell culture are mature and perfect, and the separation and culture technology of oocytes from the ovary of a female poultry has also been established. The oocytes isolated from the ovary of a female poultry can be cultured to different developmental stages in vitro Therefore, more and more scholars are investing in the research on the in vitro culture process of poultry oocytes. The initial diameter of poultry oocytes isolated in vitro or cultured in vitro is usually less than 400 μm. Alcohol dehydration and xylene before paraffin embedding sections Complex procedures such as transparency can easily cause structural damage and loss of oocytes, so the conventional paraffin-embedded method widely used in ovarian tissue sections is not suitable for the production of slices of a small number or even a single oocyte. Biomicroscopy technology brings inconvenience to the morphological observation during oocyte development and the immunohistochemical detection of some physical and chemical factors

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