RE: triglycerides

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From:"Hewlett Bryan (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA> (by way of histonet)
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Any conventional fat stain such as Oil Red O will identify triglycerides.
However, if you require a specific method, then the Calcium-Lipase method of
Adams et. al. 1966 is necessary.

Tris buffer, 0.2M pH8.0....... 15.0ml
2%w/v Calcium chloride.... 10.0ml
D. water ............................ 25.0ml
Porcine pancreatic lipase   50mg
warm solution to 37C and filter before use


1 cut 2 frozen sections and rinse in water
2 incubate one, either free-floating or slide mounted section, in lipase
reagent at 37C for 3 hours
3 place both sections in 1%w/v lead nitrate for 15 mins
4 wash in several changes D. water
5 treat with 1% ammonium sulphide for 20 secs.(pheeew)
6 wash well, counterstain with mayer's hemalum and blue, mount in


Triglycerides..... brown, nuclei...... blue
 Lipolysis is confined to the water-fat interface, thus only the surface of
large droplets may be stained. The control section should be negative, a
positive reaction indicates the presence of either free fatty acids or

Ref. Bancroft and Stevens. 4th Edition p226


> ----------
> From: 	Kowsz, Kim P[]
> Sent: 	November 10, 1999 9:45 AM
> To: 	''
> Subject: 	triglycerides
> I am looking for a staining method that will identify triglycerides in
> formalin fixed frozen mouse lung.  Thank you.

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