Gynecologic Oncology Tissue Processing (10% Formalin Fixation)
Mary Mullen M.D., Rachel Abbott
Abstract
This is a description of the steps that the Washington University's Gynecologic Oncology Tissue Bank has developed to collect surgical resection biospecimens, for molecular and genomic characterization. There are multiple protocols with various preservation methods and targeted assays. Here only the Formalin-Fixed Paraffin-Embedded (FFPE) related information is provided.
Steps
Specimen Collection & Handling (10% Formalin Fixation)
Tissue should be no bigger than 1x1x0.5 cm.
Label cassettes using pencil with Tissue Bank ID, date, and tissue type.
Place tissue in a cassette that is adequate for the size of tissue being fixated.
Place cassette in specimen cup filled with ~60 mL 10% formalin to transport to the lab at room temperature.
Tissue Processing (10% Formalin Fixation)
Tissue should be no bigger than 1x1x0.5 cm
Label cassettes using pencil with Tissue Bank ID, date, and tissue type.
Place tissue in a cassette that is adequate for the size of tissue being fixated.
The volume of formalin should be 30 times the volume of the tissue samples. Containers should be capped and leak-proof.
Leave tissue in the container for at least 24 hours in the hood.
a. For omentum samples, leave tissue for 24-48 hours depending on thickness.
b. For samples collected on Friday, leave samples in formalin over the weekend.
After fixation, transfer tissue to water rinses, 3 rinses for 15 minutes (on a shaker preferably). The volume of DI water should be 30 times the volume of the tissue sample.
After water rinses, transfer tissue to the following for the given times:
a. 30% ethanol for 30 min
b. 50% ethanol for 30 min
c. 70% ethanol for 30 min
The volume of ethanol used for these rinses should be 30 times the volume of the tissue sample.
All specimens are to be received by HistoCore in 70% ethanol, unless otherwise stated by the Histotechnologist. Samples should be turned in to HistoCore before 3:00 p.m. every day , except Fridays.
One quality control slide per sample should be labeled with Tissue Bank ID, date, and tissue type.
Complete the submission form: Histocore Submission Form.
Deliver the tissue in 70% ethanol, labeled slides, and submission form to Histocore.
When the FFPE block and slide are received back from Histocore, attach pre-printed Open Specimen labels and store in designated location.
The H&E slide should be submitted to Dr. Hagemann for QC when possible.
Research Specimen Disposal Procedure (10% Formalin Fixation)
Dispose transport container in a biohazard waste box.
Collect formalin and washes in a flask using an aspirator.
Transfer liquid collected in flask to the appropriate unwanted material container.
