Lipid staining of primary cells by Sudan red III staining method

Summary

This experiment is mainly used to identify the lipid content of cells or to identify lipid cells.

Operation method

Lipid staining of primary cells by Sudan red III staining method

Principle

The solubility of Sudan red dye in lipids is greater than its solubility in solvents. If the dye comes into contact with a specimen containing lipids, a large amount of the dye enters into the structures of the lipids, giving these structures a red color. Most of the Sudan red dye, oil red O, Nile blue and other dyes soluble in lipids staining, so that the lipid color. Can also use osmium tetroxide staining, fatty acids or choline can make osmium tetroxide reduced to osmium dioxide and black.

Materials and Instruments

Cell samples
Sudan red III 70% ethanol Formaldehyde CaCl2 Gelatin Distilled water Glycerol Phenol
Coverslips Water bath Slides Microscope

Move

I. Preparation of reagents:

1. Sudan red Ⅲ staining solution: dissolve 1g Sudan red Ⅲ in warmed 70% ethanol solution and filter for spare;

2. calcium formaldehyde fixative: mix 10ml formaldehyde and 1g CaCl2 with water to 100ml;

3. Glycerol gelatin: add 12g gelatin into 120ml distilled water in a water bath heating so that it is completely dissolved, then add 60ml glycerol and 1g phenol, mix thoroughly that is obtained. Store at 4℃ for spare;

II.Staining steps:

1. rinse the coverslip with Hanks solution for 3 times;

2. Fix with calcium formaldehyde fixative for 20min;

3. rinse the coverslips with distilled water for 3 times. 4;

4. rinse the coverslips with 70% ethanol for 3 times. 5;

5. cover the coverslips with Sudan Red III staining solution for 10-30min. 6. rinse the coverslips with 70% ethanol;

6. rinse the coverslips 3 times with 70% ethanol; 7. seal the coverslips with glycerol gelatin;

7. seal the slide with glycerol gelatin and observe;

III. Results:

The lipid-containing areas of primary cells were orange-red in color;

Caveat

When dyeing, make sure you take the time to prevent incomplete dyeing and colors that are not obvious.

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