We used to do overnight in our DAKO Autostainer many years ago, and we kept
many containers of dH20 in the incubator to make a humid chamber. Seemed to
work well - although we realized overnight was overkill and 1-2 h was
sufficient. In fact, given this we started to always put humid chambers
Hi Naira,
We routinely do a 2 hour incubation in primary, but we make sure that more
antibody is on the slide than would be necessary for most other reagents
(500-600 ul). We have not had any issues of drying. If you are concerned
about it, just make it 2 antibody steps instead of 1 on the
Yea I don't do incubations for longer than one hour on the autostainer and I
do put thin trays of water under the slides and on the sides of the machine
(it is very dry in Colorado), but I can think of a way you could do longer
incubations if you were around, put a coverslip or parafilm over the
Dear Histonetters,
Thanks everyone for several good advices.
I'll try all suggestions: increase volume from 600ul to 800ul, add more primary
antibody step and put hot water dishes inside of machine to keep humidity as
high as possible.
Hopefully it will work...
Many thanks,
Naira
Hi Histonetters!
I have a stupid question but I Have to ask.
Does anyone perform an Incubation of Ab that required more then 1 hour (2-3
hours) in autostainer? Does autostainer keep slides wet or slides sometimes are
getting dry?
I know that slides should not be dry in any step of IHC.
We have done up to an hour but not longer, if I were you I would have
two antibody steps and just to add some more reagent after an hour or
1.5 hours
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949
fax (303)
