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A rapid single cell sorting verification method using plate‐based image cytometry
Author(s) -
Zigon Eric S.,
Purseglove Suzanne M.,
Toxavidis Vasilis,
Rice William,
Tigges John,
Chan Leo LiYing
Publication year - 2018
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.23520
Subject(s) - sorting , computer science , cell sorting , flow cytometry , cytometry , microscope , single cell analysis , cell , microbiology and biotechnology , chemistry , biology , optics , physics , algorithm , biochemistry
Single cell sorting is commonly used for ensuring monoclonality and producing homogenous target cell populations. Current single cell verification methods involve manually confirming the existence of single cells or colonies in a well using a standard light microscope. However, the manual verification method is time‐consuming and highly tedious, which prompts a need for an accurate and rapid detection method for verifying single cell sorting capability. Here, we demonstrate a rapid single cell sorting verification method using the Celigo Image Cytometer. Calcein AM‐stained Jurkat cells and fluorescent beads are sorted into 96‐well half area microplates using the MoFlo Astrios EQ. Whole well bright field and fluorescent images are acquired and analyzed using the image cytometer in less than 8 min. The proposed single cell verification detection method in multi‐well microplates can allow for quick optimization of FACS instruments at flow core laboratories, as well as improvement of downstream biological assays by accurately confirming the presence of single cells in each well.