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Label‐Free Metabolic Classification of Single Cells in Droplets Using the Phasor Approach to Fluorescence Lifetime Imaging Microscopy
Author(s) -
Ma Ning,
Kamalakshakurup Gopakumar,
Aghaamoo Mohammad,
Lee Abraham P.,
Digman Michelle A.
Publication year - 2019
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.23673
Subject(s) - fluorescence microscope , microscopy , fluorescence , phasor , fluorescence lifetime imaging microscopy , biophysics , chemistry , nanotechnology , materials science , biology , physics , optics , quantum mechanics , power (physics) , electric power system
Characterization of single cell metabolism is imperative for understanding subcellular functional and biochemical changes associated with healthy tissue development and the progression of numerous diseases. However, single‐cell analysis often requires the use of fluorescent tags and cell lysis followed by genomic profiling to identify the cellular heterogeneity. Identifying individual cells in a noninvasive and label‐free manner is crucial for the detection of energy metabolism which will discriminate cell types and most importantly critical for maintaining cell viability for further analysis. Here, we have developed a robust assay using the droplet microfluidic technology together with the phasor approach to fluorescence lifetime imaging microscopy to study cell heterogeneity within and among the leukemia cell lines (K‐562 and Jurkat). We have extended these techniques to characterize metabolic differences between proliferating and quiescent cells—a critical step toward label‐free single cancer cell dormancy research. The result suggests a droplet‐based noninvasive and label‐free method to distinguish individual cells based on their metabolic states, which could be used as an upstream phenotypic platform to correlate with genomic statistics. © 2018 International Society for Advancement of Cytometry