Biomedical research paraffin section(6) Medical experimental
13, differentiation
Is to the cytoplasm with the color faded, so that the nucleus is more vivid coloring, also known as color separation. Will be sliced into 1% hydrochloric acid ethanol solution (1 part HCl +70% ethanol 100 copies) in the fade, see slice red, the color can be lighter, some a few seconds to tens of seconds. This step is key to success in HE staining, such as the improper differentiation will lead to uneven staining, or deep or shallow, stained slices obtained poor results. If the stain is moderate, cancel this step. Medical experimental
14, rinsing
Slice and then put it back into the water flowing in the blue. Low magnification microscope, see the nucleus was blue, the structure clear; cytoplasm or colorless as a standard component of connective tissue fibers. Then put into a distilled water rinse once.
15, dehydration
Slice into 50% ethanol 70% ethanol 80% ethanol in the 3 ~ 5min.
16, stained
With 0.5% ethanol solution compared eosin staining 2 ~ 5min. Eosin stained mainly the cytoplasm, coloring shading hematoxylin stained nuclei should be matched by the concentration, if the doctrinal and nuclear staining, cytoplasmic staining should also be concentrated in order to obtain contrast. Conversely, if the shallow nuclear staining, cytoplasmic staining should also be light. In eosin alcohol solution to help dropping a few drops of glacial acetic acid dye, prompting the cytoplasm easy coloring, and when dehydrated by the ethanol will not fade.
17, dehydration trial
Add 95% ethanol wash away the excess in the red, and then placed in anhydrous ethanol, 3 ~ 5min. At last, dry up the excess alcohol absorbent paper.
18, transparent
Slice into xylene - equal mixture of ethanol for about 5min, then put into pure xylene Teva Pharmaceutical Industries, Ltd.each 3 ~ 5min. Xylene should keep water should be frequently changed, or anhydrous copper sulfate with gauze bag to absorb moisture into the cylinder dyeing. Sections such as xylene, white mist appears the phenomenon, indicating dehydration not entirely, to be returned to re-dehydrated ethanol, or sliced hard to microscopic examination. clinical
19, seal Tibet: Neutral gum seal
Sections were stained, dehydrated, transparent, you can use letters Tibet agents hide their seal the purpose of permanent preservation slice, easy to microscopic examination. Possession of agents commonly used in sealing a class of agents for the dry seal Tibet, such as a neutral gum, gum, etc., Canada, and the other for the wet seal Tibet and other agents, such as glycerol gelatin. If the slice is transparent by xylene, then sealed with rubber as a possession of agents, gums can be diluted with xylene to the right consistency. If the slice is directly removed from the water or aqueous solution, then the glycerin gelatin used as a sealing agent reservoir can be used for short-term stored samples.