@inproceedings{c7decc74937d4d3ba8de5683bad9a3ed,
title = "Integrated functional characterization and optical tagging for cell-type identification",
abstract = "Imaging has become one of the most important tools for categorizing neurons based on their function. However, for a cell type identified only by its pattern of activity, the process of identifying molecular markers remains laborious. We developed physiological optical tagging sequencing (PhOTseq), a technique for tagging and ex- pression profiling of cells on the basis of their functional properties. We developed a reporter combining a green calcium indicator (GCaMP) with a photoactivatable red reporter (PAmCherry). When visualizing neuronal activity in such animals, real-time analysis allowed digital selection of cells exhibiting specific activity patterns, and photoactivation was directed specifically to those cells to tag them for later harvesting and analysis. We found that PhOTseq was capable of selecting rare cell types and enriching them by nearly 100-fold. We applied PhOTseq to the challenge of mapping receptor-ligand pairings among pheromone-sensing neurons in mice, and densely mapped the cell types responsible for encoding a specific portion of the sensory world.",
keywords = "Calcium imaging, Cell types, Gene expression, Optical tagging, Real-time analysis, Single-cell sequencing",
author = "Donghoon Lee and Maiko Kume and Holy, {Timothy E.}",
note = "Funding Information: We thank Ron Yu for sharing the TetO vector and OMP-IRES-tTA mouse, Paul H. Taghert for the HEK cell line, Heide Schoknecht for mouse care, and Dae Woo Kim for developing imaging software. We thank Terra Barnes, Juntao Chen, Joseph Corbo, Xiaoyan Fu, Cody Greer, Alessandro Livi, Richard Roberts, and Manning Zhang for suggestions and comments. This work was supported by the Hope Center Viral Vectors Core and the Mouse Genetics Core at Washington University School of Medicine. Funding: This work was funded by NIH/NIDCD R01 DC005964 and DC010381 and NIH/BRAIN 1UF1NS108176. Author contributions: Maiko Kume performed in vivo imaging and photoactivation. Donghoon Lee performed all other experiments and analysis. Timothy E. Holy supervised the study and analysis. Publisher Copyright: {\textcopyright} 2021 SPIE. All rights reserved.; Integrated Sensors for Biological and Neural Sensing 2021 ; Conference date: 06-03-2021 Through 11-03-2021",
year = "2021",
doi = "10.1117/12.2583433",
language = "English",
series = "Progress in Biomedical Optics and Imaging - Proceedings of SPIE",
publisher = "SPIE",
editor = "Hooman Mohseni",
booktitle = "Integrated Sensors for Biological and Neural Sensing",
}