[Histonet] Steamer

From:"Eric Sulkosky"

Hi All

 

My laboratory has been using the rice steamer for years and has not had
any problems. I would like to ask, have you been preheating your
solutions before trying to retrieve? You mentioned that you were having
problems with TTF and Ecad. Are these Dako antibodies? We have found the
best results with retrieving, when you preheat your solutions for twenty
minutes. Also what pH of solutions are you using?

 

Our procedure for retrieving is as follows:

 

1.	Decide which pH needs to be used. In our laboratory we use High
pH 9.9 and Regular pH 6.0
2.	For the antibodies you mentioned Ecad and TTF we use the High
pH. We have much better results. 
3.	Set up your steamer, pour your Target Retrieval and preheat in
steamer for twenty minutes.
4.	For High pH put your slides in the preheated solution and set
your timer for 40minutes. Remove the slides from the steamer and leave
in your Target Retrieval. Set another clock for 20minutes and let the
solution come to room temperature. After it is room temperature,
transfer the slides into buffer and let them sit for 5 minutes. This
helps from your tissue drying out when staining.
5.	For Regular pH put your slides in the preheated solution and set
your timer for 20minutes. Remove the slides from the steamer and leave
in your Target Retrieval. Set another clock for 20minutes and let the
solution come to room temperature. After it is room temperature,
transfer the slides into buffer and let them sit for 5 minutes. This
helps from your tissue drying out when staining.
6.	Then proceed with your staining.

 

Important note: It is very important to let your slides sit in the
retrieval and come to room temperature before transferring them into
buffer or water. The cooling down is built into the retrieval process,
so I would advise not to directly transfer into the buffer right out of
the hot steamer.

 

I hope this helps.

 

Eric

 

sulkoskye@monhealthsys.org

 

 

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