[Histonet] protocol for freezing fixed rat skeletal muscle


Hi Histontters,

Thanks for the response to my question "mapping capillary perfusion pattern".
  Here I have another question need your help. I am going to perfuse rat 
hindlimb with BSA+FITC-dextran, then to locate the FITC-dextran in frozen 
muscle tissue. To prevent FITC-dextran from diffusing out of tissue when I 
mount the tissue onto a slide, will some fixation before freezing the 
tissue be helpful? Can anyone kindly share a good protocol for freezing 
fixed muscle tissue? fixative? how long the fixation? any cryoprotection? 
By the way, I wonder if anyone knows whether the FITC will bleach during 
the fixation, freezing process?  Thanks in advance.

Lei Zhang
Biochemistry, medicine school
University of Tasmania, Australia
61 03 6226 2669

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