1. U54 CRC will coordinate with GI team, to screen for colonoscopy cases where normal tissue may be available.
2. Scheduled in advance, U54 CRC or BRPC team member will approach patient’s in-person for
consent.
3. If patient/parents’ consent, U54 CRC will coordinate with the GI Physician on procedure timing and biopsy procurement.
4. Upon day of procedure, Receive maximum 8 forceps biopsy fragments, 3-4 from the mid-ascending colon (“right colon”) and 3-4 from the mid-descending colon (or the sigmoid colon as alternate site) (“left colon”).
5. Process the biopsy fragments into the following samples. Keep samples from right colon
and left colon separate.
a. FFPE – 1 biopsy per block
b. Snap frozen:
i. 1 snap freeze in cryovial as normal
ii. 1 will be put into freezing media
1. Wash tissue with PBS to remove the blood and fecal matter.
2. Place biopsy fragment into cryopreserved tubes with 1mL freezing media.
3. Cells should be frozen slowly at 1°C/min. Put tubes in the freezing container (e.g.,
Mr. Frosty) with isopropyl alcohol and place them overnight in a -80'C freezer.
4. Next day, transfer tubes to liquid nitrogen.
6. Route FFPE samples to Research Histology Lab to:
a. Create an H&E from each FFPE sample and route to pathologist for review.
b. After review, create 2 tissue scrolls from the tissue blocks and coordinate RNA and DNA extraction.
c. Once RNA and DNA extraction is complete, the frozen LN2 samples will be shipped to lab for
scRNAseq