I would like to stain slides prepared from biopsies of GI tract for cholesterol (and cholesterol crystals), would you recommend any filipin staining protocol ?
Hi Michal,
There is a filipin (compound similar to digitoxin) method for free cholesterol.
Fixation and sections: cryostat sections, post-fixed; frozen sections of fixed tissue.
Stock solution: 2.5 mg filipin in 1 ml dimethylformamide.
Staining solution: 0.2 ml stock solution in 10 ml PBS.
Method: wash sections in PBS.
Stain for 30 min in filipin solution.
Wash in PBS (x2).
Mount in PBS or glycerine jelly.
Examine by fluorescence microscopy (excitation BG12 with 512 nam barrier filter).
Results: Free cholesterol shows strong silvery fluorescence.
original method by Kruth & Vaughan 1980 - quoted in Bancroft & Stephens histology manual.
Good luck with your project.
Thank you very much Maria. Have you got any experience in staining for cholesterol crystals in tissues?
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