Feb 24, 2020
HuBMAP: Paraffin Embedding Tissue Samples
- 1University of Florida
- Human BioMolecular Atlas Program (HuBMAP) Method Development CommunityTech. support email: Jeff.spraggins@vanderbilt.edu
Protocol Citation: Marda Jorgensen, Jerelyn Nick 2020. HuBMAP: Paraffin Embedding Tissue Samples . protocols.io https://dx.doi.org/10.17504/protocols.io.bam9ic96
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: December 18, 2019
Last Modified: February 24, 2020
Protocol Integer ID: 31137
Keywords: hubmap, embedding, tissue, histology, histpathology
Abstract
The purpose of this Standard Operating Procedure (SOP) is to outline procedures for the paraffin embedding of HuBMAP specimens.
Guidelines
- Managers and supervisors - are responsible for making sure that technicians are properly trained and equipment and facility are maintained in good working order.
- Laboratory personnel - are responsible for reading and understanding this SOP and related documents and to perform these tasks in accordance with the SOPs.
Materials
MATERIALS
ParaplastCatalog #39501006
Gauze 4x4 Non-Sterile SquaresFisher ScientificCatalog #MSD-1400249
Disposable Embedding MoldsFisher ScientificCatalog #22363553
Electron Microscopy Sciences - Tissue Embedding CenterFisher ScientificCatalog #50-293-24
Curved Medium Point General Purpose ForcepsFisher ScientificCatalog #16-100-110
General-Purpose Broad-Tipped ForcepsFisher ScientificCatalog #10-300
-Tissue Embedding Center OR Paraffin Wax Dispenser and Cold Plate
- Paraplast
-4x4 Gauze
- Preference of Forceps / Tweezers
Safety warnings
- Use physical safety precautions when working with sharps (disposable blades).
Before start
- Embedding can be a messy process, to protect your clothes it is best to wear a lab coat or apron.
- Gloves are required when working with human specimens.
Post-Processing Tissue Acquisition
Post-Processing Tissue Acquisition
Place processed cassettes in wax holding chamber of embedding center.
Stock station with embedding utensils, gauze, and disposable embedding molds.
3m 30s
Preheat embedding utensils in the forceps warming chambers located on the sides of the paraffin dispensing nozzle.
Turn on station cold plate module. .
3m 30s
Open the cassette carefully maintaining tissue orientation. Dispose of cassette lid.
30s
View the tissue sample and select an appropriately sized disposable embedding mold.
30s
Using forceps, tap the dispenser plate on the embedding center's dispensing nozzle and partially fill a mold with molten paraffin.
30s
Using heated forceps, carefully transfer the tissue inside the cassette into the mold, placing the tissue cut side down (surface viewed when opening the cassette).
Transfer the mold to the cold pad in front of the dispensing nozzle, and gently press tissue flat with forceps or a tamper.
Allow paraffin to solidify in a thin layer, to hold the tissue in position.
Once oriented with the tissue specimen in the center of the mold, place the lower basket of the emptied cassette on top of the mold.
Add additional molten paraffin to 2/3 fill the cassette basket.
Remove any bubbles that may form between the mold and the cassette by slowly lifting and repositioning the cassette.
Carefully transfer paraffin filled mold to freezing/cold plate module of embedding station.
Allow paraffin to completely solidify.
If you have additional tissue samples to embed, return to step 4 and repeat through step 11.
go to step #3 if you have additional samples.