[Histonet] Humidity chamber

[Donna Harclerode]

Thermo Electron makes a great system with a humidity chamber and a
coverplate that I have used for many years.  The slides are held with
the plastic coverplate(the coverplates are suppose to be disposable, but
I rinse them with only DI water and reuse them many times) I use 100ul
of antibody (primary, secondary- etc, normally, but have managed to use
as little as 90ul) to cover almost the whole slide.  For buffer wash I
fill up the top of the chamber with a squeeze bottle of PBS and allow to
run through in about 5 minutes. I load the slides add primary, wash,
secondary etc and only take them out for chromagen staining.
The racks worked great when I had the positive control at the top of the
slide and the test below. I do fluorescence secondaries in the
coverplates, but I not do DAB in the coverplates, but flat on the
counter.  
For HRP frozens I use the Dako endogenous peroxidase block in the
Sequenza racks. I also use the Dako avidin biotin block in the
coverplates when necessary. 
I usually incubate primary overnight in the fridge and the chambers stay
humid for at least 48 hours.  It takes a bit of practice to load the
plates, but it is so worth it.

Sequenza Racks - # 73310017 (Holds 10 coverplate assemblies)

 Coverplates -  25/pk = # 72110017
                     50/pk = # 7219950
                    250/case = # 72110013
http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_19578.pdf 4th
page


Donna Harclerode, HT, (ASCP), HTL, QIHC
Scientist / Immunohistochemistry
Cytori Therapeutics
3020 Callan Rd.
San Diego, CA 92121
858-458-0900 ext 5416
dharclerode <@t> cytoritx.com