Re: [Histonet] Calretinin

From:"Anthony Reilly"

Hi Amy
You are taking all of the right precautions to keep the sections on the slide.  If it is an intermittent  problem it may be related to the type of tissue being stained.  As Calretinin is used for the diagnosis of mesothelioma you may find that it is the cases when pleura or pleural plaques are being stained that are floating off.  Pleura by it's nature of being predominantly connective tissue and having little protein content is notorious for floating during IHC.  
The only suggestions I could make to improve your capture rate are to make sure your sections are absolutely flat on the waterbath to ensure maximum attachment to the slide as pleura like other tougher tissue like cartilage can tend to cut thicker than the wax envelope in the block creating a wave like appearance to the tissue.  Secondly try drying your sections overnight at 37C.  This will slow down your TAT but beats restaining.   Thirdly, if you get really desperate investigate other methods of HIER as this is the step most likely to cause loss of tissue from the slides.  There have been a number of papers written on using lower temperatures for longer periods ie 70C for a couple of hours instead of 100C for 20-30 minutes.
Tony Reilly
Chief Scientist
Anatomical Pathology
Pathology Queensland
Princess Alexandra Hospital
Ipswich Rd, 
Woolloongabba Q 4102
Ph: 07 32402412
Fax:0732402930 ( )
>>> "Senn, Amy R"  20/02/2008 2:34 am >>>
Hi Histoland....

We are having some problems with our Calret stains.

Our patient tissue keeps floating off & we are not sure what we're doing
wrong (if anything!)

We do use charged slides.

We float the tissue out in a separate water bath with pure DI water.

We allow the slides to stand upright for 30 minutes to air dry, and then
put them in a 62 degree oven for another 30 minutes.

We are careful not to over-handle the slides at anytime, and we make
sure our hands are clean and lotion free.

Sometimes, for weeks and weeks, the tissue is right where we put it when
it comes off the stainer.  Other times, for weeks and weeks, it's gone.
And it's only the patient--the control is where's it's supposed to be.

Is there something we're missing?  Something else we can try??

Thanks so much for all your help!!

Amy S.

Histotech, Histology Laboratory

Holy Spirit Hospital

503 N. 21st Street

Camp Hill, PA  17011

(717) 763-2124

Cell: (724) 494-2237

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