May 09, 2023

TS Procure 812 - primary fixation formalin/wet tissue (TM - 013) V.2

DOI
https://dx.doi.org/10.17504/protocols.io.bp2l61qbdvqe/v2
  • 1CSIRO ACDP
sandra.crameri
Icon indicating open access to content
QR code linking to this content
DOI: https://dx.doi.org/10.17504/protocols.io.bp2l61qbdvqe/v2
Protocol Citation: sandra.crameri 2023. TS Procure 812 - primary fixation formalin/wet tissue (TM - 013). protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l61qbdvqe/v2Version created by sandra.crameri
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: May 09, 2023
Last Modified: May 09, 2023
Protocol  Integer ID: 81613
Keywords: Procure 812, formalin, epon, primary fixation formalin, fixed histological tissue, tissues to procure, histological tissues to procure, procure 812 resin block, ts procure, tissue, wet tissue, resin block, resin, primary fixation, tm
Abstract
The method is used to process formalin fixed histological tissues to Procure 812 resin blocks.
Guidelines
Sample is not optimal for EM, best to dissect and use only the outer 2mm of tissue mass. Sample is usually derived from histology.
Protocol materials
25% Glutaraldehyde
Osmium TetroxideProSciTech
Ethanol
Propylene oxideProSciTechCatalog #C156
Propylene oxideProSciTechCatalog #C156
Propylene oxideProSciTechCatalog #C156
HEADER
SAN:
SPEC No:
OPERATOR & STEPS:
OPERATOR & STEPS:
Primary fixation
Formalin for undefined extended period of time at Room temperature

Conventional
5h 15m
2.5 % volume 25% Glutaraldehyde in 0.1 Molarity (M) Sorenson's Phosphate buffer (pH 7.2, 300mosmol/kg) for at least 00:40:00 or overnight

40m
0.1 Molarity (M) buffer 00:15:00

15m
1 % volume Osmium TetroxideProSciTech in buffer 01:00:00

1h
70 % volume Ethanol for at least 00:20:00 or overnight

20m
95 % volume Ethanol 00:20:00

20m
100 % volume Ethanol 00:20:00

20m
100 % volume Ethanol dried with molecular sieve 00:20:00


20m
50 % volume Ethanol 50 % volume Propylene oxideProSciTechCatalog #C156 00:20:00

20m
100 % volume Propylene oxideProSciTechCatalog #C156 00:20:00

20m
50 % volume Propylene oxideProSciTechCatalog #C156 50 % volume Procure 812


100 % volume Procure 81200:20:00

20m
100 % volume Procure 812 01:00:00

1h
Embed & polymerise overnight in oven at 60 °C