[Histonet] AutoStainer XL Tweaking

[sheila adey]

Hi Sally,
 
We use our Hematoxylin progressively and have no problems. Here is our protocol, hope it helps. We also use the Leica XL. You are right, I think the one dip would still be along time because of the speed of the dip.
 
Station  
              Oven    12 min
#1          Xylene   3  min
#2          Xylene   3  min
#3          100%    30 sec
#4          100%    30 sec
#5          95%      20 sec
#6          70%      20 sec
W1          H20      20 sec
#7          Gills       55 sec    III Hematoxylin (surgipath)  
W2          H20      20 sec
#11        Clarifier  20 sec     Richard Allen
W3           H20     30 sec
#12        Bluing    40 sec     Richard Allen
W4            H20    30 sec
#13         Eosin    20 sec     Surgipath
#14          95%    20 sec
#15         100%   20 sec
#16         100%   20 sec
#17        Xylene  10 sec
#18        Xylene  10 sec
#19        Xylene   Exit
 
Note that if we are bringing IHCs to water we increase the Xylene times to 5min each and the Alcohols to 2 min each. 
 
   
     
 
 
Sheila Adey HT MLTPort Huron HospitalMichigan> To: sbreeden <@t> nmda.nmsu.edu> From: mari.ann.mailhiot <@t> leica-microsystems.com> Date: Tue, 16 Oct 2007 09:27:34 -0500> Subject: Re: [Histonet] AutoStainer XL Tweaking> CC: histonet <@t> lists.utsouthwestern.edu> > Hi Sally> > I will give you a call.> > Best Regards> > Mari Ann Mailhiot BA HT ASCP> Application Specialist> Leica Technical Assistance Center> 800 248 0123 x7267> 847 236 3063 fax> mari.ann.mailhiot <@t> leica-microsystems.com> www.leica-microsystems.com> > > > "Breeden, Sara" > <sbreeden <@t> nmda.nm > su.edu> To > Sent by: <histonet <@t> lists.utsouthwestern.edu> > histonet-bounces@ cc > lists.utsouthwest > ern.edu Subject > [Histonet] AutoStainer XL Tweaking > > 10/16/2007 07:04 > AM > > > > > > > I'm setting up my Leica AutoStainer XL and would appreciate any help> with tweaking the times. I based my timing for the Autostainer on my> manual stain times but staining is not crisp and looks washed out. Docs> say eosin is fine but cellular detail is foggy. Here are my times:> > XYL - 3 at 5 min each> > 100% reagent - 2 at 30 seconds each> > 95% reagent - 1 at 30 seconds> > 80% reagent - 1 at 30 seconds> > DI water wash - 1 minute> > Hematox (currently using Sigma) - 5 minutes> > DI water wash - 1 minute> > Acid Alcohol (1%) - one dip> > DI water wash - 1 minute> > Ammonia water - 15 seconds> > DI water wash - 1 minute> > 80% reagent - 1 at 30 seconds> > 95% reagent - 1 at 30 seconds> > Eosin (alcoholic) - 10 seconds> > 95% reagent - 1 at 30 seconds> > 100% reagent - 2 at 1 minute each> > XYL - 3 at 1 minute each> > I cut my acid alcohol time down from 15 seconds to one dip but it's> still taking a lot of hematoxylin off. Should I try 0.5% acid alcohol?> I'm at a loss so far; different hematox? More time in ammonia water? If> you use this Leica AutoStainer XL, could you offer some Handy Hints? I> thank you SO much!> > > > Sally Breeden, HT(ASCP)> > NM Dept. of Agriculture> > Veterinary Diagnostic Services> > PO Box 4700> > Albuquerque, NM 87106> > 505-841-2576> > > > _______________________________________________> Histonet mailing list> Histonet <@t> lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet> > > ______________________________________________________________________> This email has been scanned by the MessageLabs Email Security System.> For more information please visit http://www.messagelabs.com/email > ______________________________________________________________________> > _______________________________________________> Histonet mailing list> Histonet <@t> lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_________________________________________________________________
Have fun while connecting on Messenger! Click here to learn more.
http://entertainment.sympatico.msn.ca/WindowsLiveMessenger