Lipid Staining

Is there a method to demonstrate lipid in botanical tissue fixed in

formaldehyde, dehydrated to 70% ETOH, embedded in LR White and cold

cured.

David Garrett "DGARRETT@GAB.UNT.EDU"

University of North Texas

Dept. Biological Sciences

(817)565-3964 Fax (817)565-4136

DGARRETT@gab.unt.edu

At the LM level there is as long as there is lipid remaining in the

tissue. You can demonstrate

it using several lipid dyes. Nile Red is my dye of choice at the

moment. It penetrates into all of the tissue but only flouresces in a

neutral environment (i.e. neutral lipids). Stain sections for 1-5 minutes

and then view using a flourescein filter pack on a flourescent microscope.

If the lipid has been dissolved out then of course there is no way to

detect it. One method for maintaining lipid in tissue is to complex it so

it is not removed. We have found the OTAP method particularly good for

this. Check out papers by John Guyton and Keith Klemp for details.

I hope this is useful.

Jay Jerome

* aka: W. Gray Jerome

* Dept. of Pathology

* Bowman Gray School of Medicine of Wake Forest University

* Medical Center Blvd

* Winston-Salem, NC 27157-1092

* 910-716-4972

* jjerome@bgsm.edu

Every EM user knows that high atomic number atoms can be used to increase

contrast and therefore "stain". Lower atomic number atoms when packed into

large molecules are also denser and can be used as "stains" in EM. That is

true for quite a number of stains, including the common lipid stain Sudan

Black B.

This particular stain will in fact show as completely opaque in TEM.

Also the opposite, the loss of lipids can be demonstrated by using a

lipase to digest these. Use gold grids if incubating the sections on the grid.

Jim Darley

Probing & Structure

Microscopy Supplies & Accessories

Phone +61 77 740 370 Fax: +61 77 892 313

A great microscopy site http://www.ultra.net.au/~pns/

pns@mailhost.ultra.net.au