Calcium staining assay
Summary
Calcium is deposited in bone tissue or pathologically altered tissue, most often in calcified tissue such as tissue necrosis, atherosclerosis and tumors. In the presence of chymotrypsin, colored chelate (calcium) components can be formed with calcium. Content source "Immunohistochemistry Experimental Techniques and Applications
Operation method
basic program
Principle
Calcium, i.e., calcified material, can be formed in a variety of tissues, and such calcium salt deposits, which may also be referred to as calcium salt deposits, have a diverse morphology, ranging from granular to lamellar and so on. Their main components are calcium phosphate and calcium carbonate. It is often stained with Dahl-McGec-Russell alizarin red S method.
Materials and Instruments
Paraffin tissue sections
Xylene Anhydrous ethanol Neutral gum Distilled water Silymarin red Acetic acid Light green Ammonium hydroxide
Dropper
Move
Alizarin Red S Staining Solution: 1 g of alizarin red S, 90 ml of distilled water, 10 ml of 1% ammonium hydroxide; pH value of this solution should be around 6.3, and the solution can be stored for at least 1 month.
Acetic acid solution: 0.5 ml of acetic acid, 100 ml of distilled water.
Brilliant green solution: 0.2 g of light green, 100 ml of distilled water.
1. paraffin tissue sections, routinely dewaxed to water. 2.
2. Stain with Alizarin Red S Stain for 5 min. 3.
3. Quickly wash with distilled water twice. 4.
4. bright green staining solution for 30 s. 5.
5. Acetic acid rapid washing twice. 6.
6. Quick dehydration with anhydrous ethanol. 7.
7. Clear in xylene and seal with neutral gum.
Caveat
1. Alizarin red staining solution should not be stored for a long time to avoid affecting the staining effect.
2. After the dyeing action, anhydrous ethanol dehydration and timely drying with filter paper, then transparent and sealing.
Common Problems
RESULTS: The calcareous material is reddish and the background is light green.
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