[Histonet] nile red staining-phospholipidosis-problems-reg

PKamalavenkatesh <@t> wockhardtin.com PKamalavenkatesh <@t> wockhardtin.com
Wed Aug 16 22:09:55 CDT 2006

Dear Histonetters,
                              This   is   with   regard   to  detection  of

phospholipidosis using nile red staining by fluorescence microscopy. I have

seen  a  number  of  discussions regarding this in our archive. Actually we

have induced phospholipidosis in rats using azithromycin since this drug is

known for inducing systemic phospholipidosis. We have taken cryosections of

liver  (10 microns thickness) and stain the section with a drop of nile red

(Sigma) initially dissolved in acetone and then diluted with Glycerol water

mixture (75:25). The final concentration of nile red is 1 microgram per ml.

We  are  able  to get two types of fluorescence one yellowish gold (neutral

lipids) and the other orange red (phospholipids). The reference is Brown et

al.,    Nile   red   staining   of   lysosomal   phospholipid   inclusions,

Histochemistry, 97: 349-354.

Now the problems I am facing are

1.Since  the  staining is done with glycerol water mixture, we are not able

      to  make  the permanent preparations and also not able to examine the

      slides under oil immersion.

2.If  there  is  any  other  protocol  that does not involve glycerol water

      mixture.  Hence  we can stain the sections with neutral red, wash the

      sections and then mount the sections.

3.Anybody can suggest me an aqueous mounting medium that will aid in making

      permanent preparations.

I also want to know if anybody is encountering the same problems


E.mail: PKamalavenkatesh <@t> wockhardtin.com

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