Jordan:  Modified Kluveer-Barrera Method for LFB

Cut 5 micron sections, dry, bake at 60 degrees C for 30 minutes.

Deparaffinize and hydrate to 95% ETOH (note not to DI H2O).

Place slides in a dish with 400 ml of 0.1% LFB (diluted in 95% ETOH).

Microwave on high power for 2 minutes (liquid should be near boiling)
(Alternatively you can put slides in a coplin jar with 50 ml of LFB and microwave for 30-40 seconds.)

Allow slides to cool in LFB at room temp for 20 minutes.

Rinse slides briefly in 95% ETOH to remove excess stain.

Rinse slides thoroughly in DI H2O.

Differentiate slides in 0.1% Lithium Carbonate for 30 seconds.

Three rinses to complete differentiation with 70% ETOH.

Three rinses with DI H2O.

Check microscopically for bright blue myelin fibers while vessel interiors are nearly colorless, RBC's should be colorless, gray matter should be almost colorless.

Repeat differentiation steps (95% ETOH and Lithium Carbonate) if further differentiation is needed.

Counterstain with routine H+E stain or Nuclear Fast Red.


Good luck!

Joe Galbraith

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan
Brod
Sent: Tuesday, March 09, 2004 10:08 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Speed Up Luxol Fast Blue Stain?


I have a doctor who needs a LFB stain today and the procedure I use requires that slides be left in the Luxol fast blue overnight.  Is there a way to speed that step up?

Have a great day!!!!

********************************************************************
Jordan W. Brod, HTL (ASCP)
Diagnostic Supervisor - Pathology
Texas Veterinary Medical Diagnostic Laboratory (TVMDL)
jbrod@tvmdl.tamu.edu


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