May 09, 2023
Version 2
TS Spurr's - primary fixation Karnovsky's - tissue (TM - 013) V.2
- 1CSIRO ACDP
Protocol Citation: sandra.crameri 2023. TS Spurr's - primary fixation Karnovsky's - tissue (TM - 013). protocols.io https://dx.doi.org/10.17504/protocols.io.8epv5j45jl1b/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: 81630
Abstract
This method is used for conventional processing of tissue to Spurr's resin.
Guidelines
All time are minimum times, it is acceptable to go over time specified for any given step. Good place steps to leave overnight or at 70% ethanol and 50/50 Spurrs/Ethanol mix.
Protocol materials
Spurrs resin mixProSciTech
In 3 steps
25% Glutaraldehyde
Step 2
16% ParaformaldehydeProSciTechCatalog #C004
Step 2
Osmium TetroxideProSciTech
Step 4
Ethanol
In 5 steps
HEADER
HEADER
SAN:
SPEC No:
OPERATOR & STEPS:
OPERATOR & STEPS:
CONVENTIONAL
CONVENTIONAL
5h 15m
5h 15m
2.5 % volume 25% Glutaraldehyde plus 4 % volume 16% ParaformaldehydeProSciTechCatalog #C004 in 0.1 Molarity (M) for at least 00:40:00
40m
Wash 0.1 Molarity (M) Sorenson's Phosphate Buffer 07.2 (300mosmol/kg) for 00:15:00
15m
1 % volume Osmium TetroxideProSciTech in buffer for 01:00:00
1h
70 % volume Ethanol for at least 00:20:00
20m
95 % volume Ethanol for 00:20:00
20m
100 % volume Ethanol for 00:20:00
20m
100 % volume Ethanol dried molecular sieved for 00:20:00
20m
50 % volume Ethanol 50 % volume Spurrs resin mixProSciTech for 00:30:00
30m
100 % volume Spurrs resin mixProSciTech 00:30:00
30m
100 % volume Spurrs resin mixProSciTech 01:00:00
1h
Oven polymerise overnight at 65 °C