May 27, 2024

Public workspaceTissue processing and freezing after surgery V.4

This protocol is a draft, published without a DOI.
  • 1Charité
Bettina Ergün
Open access
Protocol Citation: Bettina Ergün 2024. Tissue processing and freezing after surgery. protocols.io https://protocols.io/view/tissue-processing-and-freezing-after-surgery-dein3cdeVersion created by Bettina Ergün
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 18, 2024
Last Modified: May 27, 2024
Protocol Integer ID: 100654
Abstract
The aim of this protocol is to document the processing of fresh tissue after surgery.
The collected tissue must also be documented in the files which are linked down below
The protocol descibes in detail how to process the resulting tissue from following protocols:

Gewebesammlung Frischgewebe Zystektomie
Gewebesammlung Frischgewebe Prostatektomie
Gewebesammlung Frischgewebe Nephrektomie

in the current version.






Materials
Transport medium
MEM : Prozent: 98 %
Zellshield : Prozent: 2 %

MEM mit Earle's Salzen, mit 2,2 g/l NaHCO3, mit stabilem Glutamin Hersteller/Lieferant:Biosell Katalog #:BS.FG0325
ZellShield Hersteller/Lieferant:Minerva BioLabs Katalog #:13-0050



Freezing medium
FKS : Prozent: 90 %
DMSO : Prozent: 10 %

FBS Superior stabil Hersteller/Lieferant:Biosell Katalog #:FBS. S 0615

sterile forceps, scapels and petri dishes
1 ml Cryotubes

Sample processing in the lab
Sample processing in the lab
Fresh tissue should be processed on the same day if possible, but can be stored at 4°C for a maximum of overnight.
To reduce pathogens, the tissue should be incubated for Duration01:00:00 atTemperatureRoom temperature

1h
Patient#OrganVisit


Note
Comments/remarks on the tissue/processing:

The tissue is divided in the lab into pieces with an edge length of 1-2 mm.
Mix the fragments of a tumor piece so that each aliquot is as representative as possible of the tumor area.
16 (decision from June 29, 2023; previously: 8) fragments are stored in 1 ml of freezing medium in a cryotube.
If there are enough fragments for more than 4 cryotubes, more fragments can be combined in one tube. A maximum of 5 cryotubes of each entity are frozen.
New procedure for storage and dokumentation started in May 2024 (change of storage tubes to LVL-Tubes with Barcode)
scan tube numbers and fill in the list mentioned in Step 9

Cryo tubes should be additionally labelled as follows:
Patient ID, Organ, T1/T2 resp. N1/N2...
No. of Pieces
Date
Freeze samples in Mr. Frosty overnight.
On the next day transfer the tubes to N2 in the right order and document the location in the N2 binder and the list mentioned in Step 9
Documentation
Documentation
Please add pictures of:
  • the orgin of the sample in the organ (if avaible)
  • each tissue piece that has been taken before cutting the fragments. The picture should include the tissue ID (in the picture)
  • the documentation sheet


Note