[Histonet] frozen in mouse tissues

[Blanca Lopez]

Hello Histonets,
I am looking to solve this issue:

what we usually do for froze section is embedded tissue in OCT and then do the sectioning. It works well for other tissues. However, because lung is hollow inside, when I used the same approach (just embed in OCT), the entire lung collapse during sectioning and I cannot even get tissue slides. I checked online and other people say I need to perfuse lung with PBS:OCT 1:1 buffer before embedded into OCT, so the lung will be filled by OCT. I tried this method, and finally I was able to get lung sliced. However, the morphology is bad. Please check the figure below. The lung tissue sections stained with hematoxylin alone (the spleen sections are fine, it's stained with anti-CD4 and hematoxylin). I heard some people also say a prior fixation by formalin before OCT embedding will help to get better morphology

Can somebody help?
Thanks

Blanca Lopez
Histotech (ASCP)
UTSW Tissue Resource K1.210
Simmons Comprehensive Cancer Center
UT Southwestern Medical Center
Telephone: 214-648-7598
Email: Blanca.Lopez at utsouthwestern.edu


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