HE stained paraffin sections the basic process of making
HE stained paraffin sections the basic process of making
1, drawn and fixed:
Fresh corpses from a human or animal tissue removed (usually the thickness of no more than 0.5 cm) into a pre-allocated in a good fixative (10% formalin, Bouin's fixative, etc.) to enable the organization of cells and protein denaturation solidification, in order to prevent cell death of the autolysis or bacterial decomposition, so as to maintain the original cell morphology. pharmaceutical CRO
2, dehydration transparent:
Tends to be used from low concentrations to high concentrations of ethanol as dehydrating agent, and gradually take off tissue mass in the water. Tissue and then placed in both soluble in alcohol, then dissolved in xylene in paraffin and transparent agent transparent to replace xylene alcohol block out the organization in order to dip wax embedding.
3, the Baptist wax embedding:
Will have been placed in a transparent tissue mass has been melted paraffin wax, the wax into the melting tank insulation. To be completely immersed in paraffin-embedded tissue after: first preparation of a good container (such as the folding of a small carton), pour the wax has been melted and rapidly take folder already soaked tissue into which paraffin wax. Serve cooled solidified into blocks. Good embedded tissue hardens, the slicing machine can be cut into thin slices. (Biological experiments show channel Vital Helesys Pvt. Ltd. )
4, slice and patch:
Would be a good wax-embedded blocks fixed on the slicing machine, cut into thin slices, usually 5-8 microns thick. Folds are often cut in thin slices, to put the water heating ironing, paste to the glass slide and let dry in 45 incubator.
5, dewaxing staining:
HE stain commonly used to increase the tissue cell structure of the various parts of the color difference, benefit observed. Hematoxylin (Hematoxylin, H) is a basic dye, can be the nucleus and the cell ribosome dyed blue-violet, is the structure of basic dye staining basophilic. Eosin (Eosin, E) is an acidic dye, can the cytoplasm stained red or light red, is the structure of acid dyes eosinophilic. Staining is required before taking off with xylene in the paraffin sections, and then by the high concentration to low concentration of alcohol, and finally into the distilled water, can be dyed.
HE staining process is:
would have been put into the distilled water after the slicing hematoxylin staining in aqueous solution a few minutes.
acidic water and ammonia in the color separation, each a few seconds.
running water into distilled water after 1 hour and moment.
into the 70% and 90% alcohol in the dehydration of 10 minutes.
into the alcohol-Eosin Staining staining 2-3 minutes.
6, dehydration and transparent:
After staining sections were pure alcohol dehydration, and then by xylene to make a transparent slices.
7, Sealing:
Will have a transparent slice on the Canadian gum drops, covered with coverslip Sealing. To be a little dry gum, paste standard, sliced specimen can be used.