[Histonet] Greetings and IMHC Questions

Lewis, Patrick patrick.lewis <@t> seattlechildrens.org
Mon Aug 23 14:03:37 CDT 2010

Hello everyone,


My name is Patrick Lewis and I just started working for Children's Hospital in their Research Department.


It's been a long time since I've done any Immunohistochemistry, (~10 years), but in this new job, I will be doing a lot of it with both paraffin and frozen sections that I will make myself.


Anyway, I am about to try staining my first frozen slides and I have a couple of quick questions.  


 I cut some practice slides, human tonsil, and have stored them at -80 in a slide box.

How much longevity will they have?


I'd like to be able to cut slides and store them unfixed at -80 for up to one month until I use them


I know its better to use them the same day, but it would be nice to be able to cut them on one day and use them as needed


Also, is there a good place to find general IMHC procedures?   It's been a long time and I want to re-familiarize myself with some general protocols



I am using Superfrost Plus slides, but we may have some problems with the sections falling off the slide.  Does anyone have any suggestions on methods to avoid tissue loss from the slide?



Also, I am freezing my tissues in OCT in cryomolds that are lowered into an iso-pentane filled  ½ coke can which is immersed in a bucket of dry-ice for 5 minutes.  .  I wanted to avoid using liquid nitrogen and am hoping that with the dry ice method I would still get good freezing of the block.  Are there any recommendations for OCT freezing?


Thanks for all your help.





Patrick Lewis

Research Associate II-Bench| Infections and Prematurity

Seattle Children's Research Institute

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patrick.lewis <@t> seattlechildrens.org

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