Re: [Histonet] IF troubleshooting

From:"Gudrun Lang"

RE: [Histonet] IF troubleshootingTeresa,
the doctors complain about a misty look of the slides. They believe something like too little deparaffination. There is some reaction, but not clear. I have to ask them about further informations.
We did the same procedure like everytime. (this time even with very fresh xylol).
We had this problem once before a while. We repeated the whole procedure on the same slides incl. dryer and xylol. Then the doctors were satisfied, but I have my worries about the significance of the test. (Besides every antibody was negativ)  We don't run controls.

our protocol:
30 min 60 degree dryer
3 x 7 min Xylol
each 2 min Alcohol 96-80-50
15 min 0,1% Protease in PBS 37 degree
cool in cold PBS
30 min fitc-labeled antibodies
3 x 5 min rinse with PBS
mounting with IF-medium

Can you help us?

  ----- Original Message ----- 
  From: Flores, Teresa 
  To: Gudrun Lang 
  Sent: Monday, August 30, 2004 6:14 PM
  Subject: RE: [Histonet] IF troubleshooting

  Gudrum, what kind of trouble are you having? 

  -----Original Message----- 
  From: [] On Behalf Of Gudrun Lang

  Sent: Monday, August 30, 2004 9:54 AM 
  To: Histonetliste 
  Subject: [Histonet] IF troubleshooting 

  Can anyone give me a website with IF-troubleshooting, especially on FFPE renal biopsies? 
  Or any other information in electronical form. 

  thanks in advance 
  Gudrun Lang 
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