[Histonet] Indirect Immunofluorescence

[Flores, Teresa]

Juanita,
Our EM Lab performs immunofluorescent studies on renal, oral and skin
biopsies. 
We supply our "customers" with Michel's Transport Media (sold commercially
as Zeus) 
We use "charged" slides to pick up all our frozen sections (sectioned at 6
microns). 
We fix the frozen oral, skin or renal sections in EM Grade cold Acetone for
30 minutes, PBS for 30 min and then incubate with titrated antibodies
(according to manufactured are titrated): IgG, IgM, IgA, C3, C1q, C4d(C4d is
time consuming, costly and requires 4 steps=2hrs), CMV, Polymiovirus
(requires 2 steps = 2 hrs), Kappa, Lambda, and Fibrinogen for 1 hour.
PBS 2x washes
Coverslip with BioMedia
Hold in refrigerator without a bulb and no fixative inside the refrigerator
until pathologist is ready to view. 
NOTE: no immunofluorescent light is allowed on sections as they tend to
fade.
Our products are most but not exclusively from DAKO.
Teresa Flores
LSUHSC
New Orleans, LA
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Brody,Juanita X
Sent: Wednesday, July 21, 2004 4:57 PM
To: 'histonet <@t> pathology.swmed.edu'
Subject: [Histonet] Indirect Immunofluorescence

 We do indirect IF on skin specimens and  having problems with false
positives and background staining. Is there any one doing this procedure who
would be willing to share the source of money slides, control sera, and
procedure?
Thanks in advance.
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