Submitting tissue for frozen sections
Size of specimens:
- Tissue should be trimmed to specified region before bringing to lab.
- Clean off all fat.
- Size to fit embedding molds.
- If cross sections are needed do not have tissue too long.
Preparation of tissue:
- Freeze fresh frozen tissue immediately in liquid Nitrogen (tubes or vials).
- If embedding is done in OCT do not put back in liquid Nitrogen.
- Frozen tissue can be stored in 80° freezer but transport to lab in liquid nitrogen or on dry ice only. DO NOT use crushed ice.
Acceptable fixative:
- 4% Paraformaldehyde (PFA) – place tissue in fixative immediately.
- Fixation times will vary. For example:
small lymph node/mouse ovary – 2-3 days
pancreas/lung/kidney/etc. – 5 days
Fix mouse testis in PFA for 2 days, cut in half and fix for 2-3 weeks or more. - We will provide 30% sucrose to cryoprotect tissue (sucrose made in PBS Buffer pH 7.2 – 7.3).
Frozen sections:
- Tissues will be cut at 5 – 6 microns
- Two sections per slide unless otherwise specified.
- Sections are placed on superfrost/plus slides (charged).
Slide boxes:
Slide boxes need to be brought to the lab with tissue or will be provided at a charge of $5.00 each.
You must bring dry ice when picking up frozen tissue and prepared slides.
You need to enter a service request in iLab (Research Histology Core) under Cryo Sections