Human Pregnant Fallopian Tube Tissue Collection and Preservation Methods - UCSD Female Reproductive TMC

Fallopian tubes should be collected into empty sterile cups right after sterilization (salpingectomy or tubal ligation). Keep left and right tubes separate.

Take a photo of the tubes prior to sampling. If pieces of the tissue were collected for clinical pathological examination, maintain the original orientation of tubes from the fimbrial (distal) end to the tubal (proximal) end after such sampling.

Wash the tissues in cold PBS while maintaining the original orientation.

Separate the fimbriated end from the tube (if not already separated).

Divide the fimbriated end lengthwise into 2 or 3 equal sized sections (depending on size).

Place one fimbriated piece into a 10% formalin-filled tube (1^st priority), the other into RNAlater (2^ndpriority), and the third to be snap-frozen (3^rdpriority).

Collect cross sections of the tube into 10% formalin (x1), RNAlater (x3) and to be snap-frozen (x3). Each piece should be about 5mm in length, depending on the total length of the tube. If there is an excess of tissue, the formalin piece(s) can be larger.

Collect the formalin-preserved piece from the most distal end (closest to the fimbria).

For the right tube , collect the RNAlater pieces from the center of the tube and the snap frozen pieces from the proximal end. T1 should be most distal, then T2, then T3 should be most proximal.

For the left tube , collect the snap frozen pieces from the center of the tube and the RNAlater pieces from the proximal end. T1 should be most distal, then T2, then T3 should be most proximal.

Drop the snap frozen labeled tubes into liquid Nitrogen. Leave for ~2-10 minutes, then store in -80C freezer.

Place RNAlater tubes into a 4C fridge. Let these sit for 24-48 hours, then remove the RNAlater with a sterile transfer pipette and store in a -80C freezer.

After one week in formalin, place each formalin-fixed piece into a labeled cassette to be processed into FFPE blocks.