[Histonet] Oil Red O staining for atherosclerosis lesions

[delphine eberle]

Hi,
 
We stain frozen aortic root sections with oil red o (diluted in propylen glycol) as follow:

  1.       Fix slides in 10% formalin (5 min)
  2.       Rinse in 1X PBS (5 min)
  3.       Dip in Distilled H2O until clear (3 dips)
  4.       Transfer slides into 100% propylene glycol (2 min)
  5.       Transfer slides into 0.5% Oil Red O stain (overnight) (Filter before use)
  6.       Differentiate slides in 85% aqueous propylene glycol (2 min)
  7.       Wash slides in distilled H2O (3 min)
  8.       Counter-stain nuclei in Hematoxylin Gil 1X (2min)
  9.       Dip in Tap H2O until clear (3 dips)
  10.   Bluing Reagent (0.1% Sodium Bicarbonate in distilled H2O) (3-4 dips)
  11.   Tap H2O (30 sec)
  12.   Coverslip and seal slides using Aquatex mounting medium
Recently, we've been getting dirty staining with multiple droplets left on the slides.
We tryied to do more washes in 85% propylene glycol but it does not seem to improve.
Would anybody have ideas to solve this?
 
Thanks!!
Delphine


Delphine Eberlé PhD 
UCSF Department of Vascular Surgery
VA Medical Center - NCIRE
Building 2 - room 410
4150 Clement Street 
San Francisco, CA 94121, USA
Tel: 415 221 4810 ext.2984
delphine.eberle <@t> ucsfmedctr.org











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