Feb 28, 2024

Public workspaceOMS Atlas OCT Spatial Mapping - Limited

 Forked from OMS Atlas OCT Spatial Mapping
DOI
https://dx.doi.org/10.17504/protocols.io.8epv5xy24g1b/v1
  • Brett Johnson1,
  • Danielle Galipeau1,
  • George Thomas2
  • 1Oregon Health & Science University;
  • 2Knight Comprehensive Cancer Institute, Oregon Health & Science University
Brett Johnson
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DOI: https://dx.doi.org/10.17504/protocols.io.8epv5xy24g1b/v1
Protocol Citation: Brett Johnson, Danielle Galipeau, George Thomas 2024. OMS Atlas OCT Spatial Mapping - Limited. protocols.io https://dx.doi.org/10.17504/protocols.io.8epv5xy24g1b/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: February 28, 2024
Last Modified: February 28, 2024
Protocol Integer ID: 95829
Keywords: oms atlas oct spatial mapping, oms atlas, oct block, specimens for downstream analysis, specimen
Abstract
This protocol describes the procedure by which the OMS Atlas serially sections an OCT block, prepares the resulting slides and samples, and then distributes the specimens for downstream analysis.
Materials
MATERIALS
ReagentSuperfrost Plus Microscope SlidesFischer ScientificCatalog #12-550-15

Additional equipment:
  • Cryostat
Troubleshooting
Before start
Transfer OCT blocks to OHSU Knight Histopathology Shared Resource (HSR) for sectioning and processing.
Preparation
Verify the identity of the OCT block to be cut against written request for sectioning.
Remove OCT block from Temperature-80 °C freezer and acclimate to cryostat (Temperature-20 °C ) for minimum of Duration03:00:00 .

Label all slides and cryotubes with a unique BEMS ID and Part#, corresponding to the written request and OCT spatial map (below).
ABCDE
Part#DescriptionThicknessAssayRecipient
1Superfrost Plus slide5µmH&EOHSU, HSR
2Superfrost Plus slide5µmCyclic Immunofluorescence (Tumor Panel)HMS, Alyce Chen
3Superfrost Plus slide5µmCyclic Immunofluorescence (Tumor Panel)HMS, Alyce Chen
4Remainder of OCT blockNASingle Cell Indexing ATAC SequencingOHSU, Andrew Adey

Sectioning
Affix OCT block to cryostat chuck.
Orient and face block to get adequate amount of core.
Note: Avoid excessive facing to reduce tissue loss.
Set cryostat to 5 micron sections.
Note: All sections cut from here on should be sequential. The serial order, adjacency, and consistent orientation of the sections are all important factors. Please note any deviations from the protocol.
Cut three sections at 5 microns (Part#1-3) and affix onto appropriately labeled slide according to OCT spatial map (step #3 above).
Place all slides and remaining OCT block in Temperature-80 °C freezer.
Note: No slides are to be fixed under this protocol.

Processing
Perform hematoxylin and eosin (H&E) staining on slide labeled Part#1 (see OCT spatial map in step #3 above).
Deliver unstained slides (Part#2 and 3) and remainder OCT block (Part#4) to BioLibrary for distribution.
Note: Keep samples frozen at all times. Store at Temperature-80 °C . Transfer/ship on dry ice.