Maria,
        Still use iso pentane but cool it in a bath containing dry ice and 
IMS. You will get the temperature down to about -70. Your freezing should 
be fine because the critical factor is to get the temperature below -20 as 
quickly as possible to avoid ice crystal formation. I have use this method 
many times and never had a failure. If you are worried try out a test 
sample first before getting on with your precious biopsies.
                                        Regards,
                                                    Tony
Histopathology Group
Asthma Biology Department.
RIRP CEDD.
GlaxoSmithKline Medicines Research Centre,
Gunnelswood Road,
STEVENAGE,
Hertfordshire.
  SG1 2NY
tel.          +44 (0)1438 764117
fax.         +44 (0)1438 764782
email.    Tony.J.Savage@gsk.com
mobile    +44 07753609835





maria.doyle@ucd.ie

24-Mar-2003 14:19

 
 

        To:     histonet

        cc: 
        Subject:        Liquid nitrogen problem

Dear all,

I am new to this work and if somebody could help me with this 
problem I would be very grateful. 
We are going to obtain skeletal muscle samples from surgery 
which we want to use for immunohistochemistry and in situ 
hybridization. We wanted to mount the muscle sample on cork in 
OCT and freeze it in isopentane/ liquid nitrogen there at the 
hospital. However there is no facility for getting liquid nitrogen at 
the hospital and transporting it there would be difficult. My question 
is would it be possible to freeze the muscle sample on dry ice, 
transport it back to the lab and then freeze in isopentane/liquid 
nitrogen? Could this sample still be used for immunohistochemistry 
and in situ hybridization?

Thanks to anyone that can help!

Maria Doyle
PhD Candidate
University College Dublin,
Ireland