PT  - JOURNAL ARTICLE
AU  - Xiaodan Zhang
AU  - Chuansheng Hu
AU  - Chen Huang
AU  - Ying Wei
AU  - Xiaowei Li
AU  - Miaomiao Hu
AU  - Hua Li
AU  - Ji Wu
AU  - Daniel M. Czajkowsky
AU  - Yan Guo
AU  - Zhifeng Shao
TI  - Robust Acquisition of High Resolution Spatial Transcriptomes from Preserved Tissues with Immunofluorescence Based Laser Capture Microdissection
AID  - 10.1101/2021.07.13.452123
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.07.13.452123
4099  - http://biorxiv.org/content/early/2021/07/14/2021.07.13.452123.short
4100  - http://biorxiv.org/content/early/2021/07/14/2021.07.13.452123.full
AB  - The functioning of tissues is fundamentally dependent upon not only the phenotypes of the constituent cells but also their spatial organization in the tissue. However, obtaining comprehensive transcriptomic data based on established phenotypes while retaining this spatial information has been challenging. Here we present a general and robust method based on immunofluorescence-guided laser capture microdissection (immuno-LCM-RNAseq) to enable acquisition of finely resolved spatial transcriptomes with as few as tens of cells from snap-frozen or RNAlater-treated tissues, overcoming the long-standing problem of significant RNA degradation during this lengthy process. The efficacy of this approach is exemplified by the characterization of differences at the transcript isoform level between cells at the tip versus the main capillary body of the mouse small intestine lacteal. With the extensive repertoire of phenotype-specific antibodies that are presently available, our method provides a powerful means by which spatially resolved cellular states can be delineated in situ with preserved tissues. Moreover, such high quality spatial transcriptomes defined by immuno-markers can be used to compare with clusters obtained from single-cell RNAseq studies of dissociated cells as well as applied to bead-based spatial transcriptomics approaches that require such information a priori for cell identification.Competing Interest StatementThe authors have declared no competing interest.