Aug 18, 2022

Public workspaceCoating superfrost microscope slides with gelatin-chromium potassium sulfate

DOI
https://dx.doi.org/10.17504/protocols.io.n2bvj65owlk5/v1
  • Benjamin Trist1,
  • Alejandra Rangel1,
  • Rain Kwan1
  • 1The University of Sydney, Sydney, Australia
Benjamin Trist
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DOI: https://dx.doi.org/10.17504/protocols.io.n2bvj65owlk5/v1
Protocol Citation: Benjamin Trist, Alejandra Rangel, Rain Kwan 2022. Coating superfrost microscope slides with gelatin-chromium potassium sulfate. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvj65owlk5/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: August 12, 2022
Last Modified: May 31, 2024
Protocol Integer ID: 68563
Keywords: Microscope slide coating, Histology, Immunohistochemistry, Tissue sections, Mounting, ASAPCRN, superfrost microscope slides with gelatin, coating superfrost microscope slide, microscope slides with gelatin, floating immunohistochemical staining, microscope slide, immunohistochemical staining, chromium potassium sulfate, chromium potassium sulfate this protocol, preparation for histology, immunohistochemical analysis of thin tissue section, gelatin, chrom alum, immunohistochemical analysis, slide, histology, tissue, tissue section, thin tissue section
Funders Acknowledgements:
Michael J Fox Foundation
Grant ID: ASAP-000497
Abstract
This protocol describes how to coat microscope slides with gelatin-chromium potassium sulfate (gelatin-chrom alum) in preparation for histology or immunohistochemical analysis of thin tissue sections. Slides coated with gelatin-chrom alum exhibit much better retention of tissues mounted after free-floating immunohistochemical staining, especially during alcohol and xylene dehydration steps immediately prior to mounting media embedding and cover slipping.
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Materials
Equipment:

  • Oven
  • Heated magnetic stirrer
  • Thermometer
  • Chemical spatulas
  • Slide racks
  • Heat resistant beaker compatible with slide racks

Consumables:

  • Magnetic stirrer bars
  • Slide storage box
  • Superfrost microscope slides

Key reagents:
  • ReagentGelatinMerck MilliporeSigma (Sigma-Aldrich)Catalog #G2500 .
  • ReagentChromium potassium sulfateMerck Millipore (EMD Millipore)Catalog #101036 .
Troubleshooting
Experimental Outline
2d 0h 0m 50s
Place heated magnetic stirrer in fume hood.
Pre-heat oven to Temperature42 °C .
Heat Amount1 L dH20 to Temperature50 °C - Temperature60 °C and completely dissolve Amount10 g gelatin with aid of magnetic stirrer.
Add Amount1 g chromium potassium sulfate - solution should turn a pale green/blue and be completely clear.
Pipetting
Once dissolved reduce temperature to Temperature40 °C -Temperature50 °C .
Place slides for coating into designated slide coating rack(s).
Dip rack of slides into warm (Temperature40 °C -Temperature50 °C ) gel mixture for approximately Duration00:00:20 -Duration00:00:30 .

50s
Shake the excess liquid from the rack.
Repeat steps 7 and 8.
Place slide rack(s) to dry in the oven at Temperature42 °C for Duration48:00:00 .
2d
Store slides in dust-free slide storage box.
Discard Gelatin-Chrom Alum solution into appropriate waste container.