May 09, 2023

Public workspaceTS Spurrs - primary fixation Karnovsky's - tissue (TM - 013)

DOI
dx.doi.org/10.17504/protocols.io.6qpvr4jypgmk/v1
  • 1CSIRO ACDP
sandra.crameri
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DOI: dx.doi.org/10.17504/protocols.io.6qpvr4jypgmk/v1
Protocol Citation: sandra.crameri 2023. TS Spurrs - primary fixation Karnovsky's - tissue (TM - 013). protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr4jypgmk/v1
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: 81634
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
Reagent25% Glutaraldehyde
Reagent16% ParaformaldehydeProSciTechCatalog #C004
ReagentOsmium TetroxideProSciTech
ReagentEthanol
ReagentEthanol
ReagentEthanol
ReagentEthanol
ReagentEthanol
ReagentSpurrs resin mixProSciTech
ReagentSpurrs resin mixProSciTech
ReagentSpurrs resin mixProSciTech
HEADER
HEADER
SAN:


SPEC No:


OPERATOR & STEPS:


OPERATOR & STEPS:
CONVENTIONAL
CONVENTIONAL
5h 15m
5h 15m
Concentration2.5 % volume Reagent25% Glutaraldehyde plus Concentration4 % volume Reagent16% ParaformaldehydeProSciTechCatalog #C004 in Concentration0.1 Molarity (M) for at least Duration00:40:00

40m
Wash Concentration0.1 Molarity (M) Sorenson's Phosphate Buffer Ph07.2 (300mosmol/kg) for Duration00:15:00

15m
Concentration1 % volume ReagentOsmium TetroxideProSciTech in buffer for Duration01:00:00

1h
Concentration70 % volume ReagentEthanol for at least Duration00:20:00

20m
Concentration95 % volume ReagentEthanol for Duration00:20:00

20m
Concentration100 % volume ReagentEthanol for Duration00:20:00

20m
Concentration100 % volume ReagentEthanol dried molecular sieved for Duration00:20:00

20m
Concentration50 % volume ReagentEthanol Concentration50 % volume ReagentSpurrs resin mixProSciTech for Duration00:30:00


30m
Concentration100 % volume ReagentSpurrs resin mixProSciTech Duration00:30:00

30m
Concentration100 % volume ReagentSpurrs resin mixProSciTech Duration01:00:00

1h
Oven polymerise overnight at Temperature65 °C