PT  - JOURNAL ARTICLE
AU  - Anna Bäckström
AU  - Laura Kugel
AU  - Christian Gnann
AU  - Hao Xu
AU  - Joseph E. Aslan
AU  - Emma Lundberg
AU  - Charlotte Stadler
TI  - A sample preparation protocol for high throughput immunofluorescence of suspension cells
AID  - 10.1101/2020.01.05.895201
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.01.05.895201
4099  - http://biorxiv.org/content/early/2020/01/06/2020.01.05.895201.short
4100  - http://biorxiv.org/content/early/2020/01/06/2020.01.05.895201.full
AB  - Imaging is a powerful approach for studying protein expression and has the advantage over other methodologies in providing spatial information in situ at single cell level. Using immunofluorescence and confocal microscopy, detailed information of subcellular distribution of proteins can be obtained. While adherent cells of different tissue origin are relatively easy to prepare for imaging applications, nonadherent cells from hematopoietic origin, present a challenge due to their poor attachment to surfaces and subsequent loss of a substantial fraction of the cells. Still, these cell types represent an important part of the human proteome and express genes that are not expressed in adherent cell types. In the era of cell mapping efforts, overcoming the challenge with suspension cells for imaging applications would enable systematic profiling of hematopoietic cells. In this work, we successfully established an immunofluorescence protocol for preparation of suspension cell lines and peripheral blood mononucleated cells (PBMC) and human platelets. The protocol is based on a multi-well plate format with automated sample preparation, allowing for robust high throughput imaging applications. In combination with confocal microscopy, the protocol enables systematic exploration of protein localization to all major subcellular structures.