10x Genomics Xenium for FFPE User Guides

Andrew Houston

Published: 2024-03-02 DOI: 10.17504/protocols.io.81wgbxjonlpk/v1

Abstract

Xenium allows for in situ transcript detection of selected probe set of RNA targets revealing spatially resolved expression patterns at nearly single-cell resolution.

Steps

Tissue processing and QC

1.

Determine suitable samples of interest by performing a test described in CG000578, sectioning guidelines described in Step 1 pgs. 18-25 and H&E staining in Appendix pgs. 38-45

CG000578_Demonstrated_Protocol_XeniumInSituProtocolsFFPE_TissuePreparationGuide_Rev_C.pdf

2.

Evaluate section and staining quality, only choose samples or regions free of detachment or artifacts

Xenium slide preparation

3.

Utilize CG000578 for section placement on Xenium slides, follow Step 3, pgs. 29-33

4.

After drying slide overnight or up to 4 weeks, proceed with deparaffinization and decrosslinking steps found in CG000580 CG000580_Demonstrated_Protocol_Xenium_FFPE_Deparaffinization_Decrosslinking_RevC.pdf

5.

Immediately following decrosslinking start to prepare probes of interest as outlined in CG000582 CG000582_XeniumInSitu_GeneExpression_UserGuide_RevE.pdf

Initiate probe hybridization overnight, continue with the protocol the next day

Xenium analyzer

6.

Prepare Xenium instrument as outlined in CG000584CG000584_Xenium_Analyzer_UserGuide_RevD.pdf

7.

Load slides and initiate sample scan

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