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Some Staining Procedures:
1. Safranin-Fast Green Method:
Keep the material to be stained in safranin for three to five minutes and then wash it with water. See under the microscope that only thick-walled cells are stained Excess of stain is destained by acid alcohol. Again wash the material very thoroughly with water so that even traces of acid are removed.
Now stain the material with few drops of fast green for few seconds. Time for keeping material in fast green varies from few seconds to one minute for different materials. Wash the material with glycerin and mount in a drop of glycerin.
With this method, all thick-walled cells get red stain and thin walled cells the green stain. It can be tabulated as follows:
Select a thin section
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Slain with safranin (for 3 to 5 minutes)
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Wash with water
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Destain with acid alcohol (if need be)
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Wash thoroughly with water to remove the traces of acid
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Slain with fast green (few seconds to one minute)
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Wash with glycerine
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Mount in glycerine
2. Safranin-Aniline Blue Method:
Follow exactly the same procedure as mentioned above except that in place of fast – green use aniline blue.
3. Haematoxylin-Safranin Method:
Keep the sections in delafield haematoxylin for tour to five minutes and remove the excess of stain with water. Wash with ammonia. Wash the material very thoroughly with water. Now stain with safranin for few minutes. Wash the sections with glycerine for removing excess of stain and mount in glycerine.
It can be tabulated as follows:
Select a section
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Stain with haematoxylin (4-5 minutes)
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Wash with water
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Wash with ammonia water till stain turns blue
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Wash with lap water
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Slain with safranin (2-3 minutes)
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Wash in glycerine.
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Mount in glycerine.