Cell Interaction Analysis by Imaging Flow Cytometry

Payés, Cristian; Rodríguez, José A.; Friend, Sherree; Helguera, Gustavo

doi:10.5772/51147

Cited by 6 publications

(2 citation statements)

References 24 publications

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“…A technical barrier preventing addressing these questions is the ability to synchronize a sufficient number of cell-cell interactions for monitoring entire signaling processes before and after cell contact. Among conventional methods adopted for studying cell signaling, flow cytometry is unable to maintain continuous contact between two immune cell types (19,28). In cell interaction assays conducted in bulk (20,21), different cell types are forced into contact by gravity or centrifugation, which renders little control on location and timing of cell contacts (29)(30)(31).…”

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confidence: 99%

Dynamic Mitochondrial Migratory Features Associated with Calcium Responses during T Cell Antigen Recognition

Raddatz

Zhou

et al. 2019

The Journal of Immunology

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A T cell clone is able to distinguish Ags in the form of peptide–MHC complexes with high specificity and sensitivity; however, how subtle differences in peptide–MHC structures translate to distinct T cell effector functions remains unknown. We hypothesized that mitochondrial positioning and associated calcium responses play an important role in T cell Ag recognition. We engineered a microfluidic system to precisely manipulate and synchronize a large number of cell–cell pairing events, which provided simultaneous real-time signaling imaging and organelle tracking with temporal precision. In addition, we developed image-derived metrics to quantify calcium response and mitochondria movement. Using myelin proteolipid altered peptide ligands and a hybridoma T cell line derived from a mouse model of experimental autoimmune encephalomyelitis, we observed that Ag potency modulates calcium response at the single-cell level. We further developed a partial least squares regression model, which highlighted mitochondrial positioning as a strong predictor of calcium response. The model revealed T cell mitochondria sharply alter direction within minutes following exposure to agonist peptide Ag, changing from accumulation at the immunological synapse to retrograde movement toward the distal end of the T cell body. By quantifying mitochondria movement as a highly dynamic process with rapidly changing phases, our result reconciles conflicting prior reports of mitochondria positioning during T cell Ag recognition. We envision applying this pipeline of methodology to study cell interactions between other immune cell types to reveal important signaling phenomenon that is inaccessible because of data-limited experimental design.

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confidence: 99%

Dynamic Mitochondrial Migratory Features Associated with Calcium Responses during T Cell Antigen Recognition

Raddatz

Zhou

et al. 2019

The Journal of Immunology

View full text Add to dashboard Cite

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“…The statistics are often limited to a pixel-by-pixel analysis of correlation using Pearson’s correlation coefficient or Mander’s overlap coefficient (Zinchuk and Zinchuk, 2008), or more advanced analysis techniques such as spatial point pattern analysis (Burguet and Andrey, 2014; Helmuth et al , 2010) or protein–protein interaction models (Johnson et al , 2015). By contrast, there is a lack of statistical techniques to study cell–cell interactions (Payés et al , 2012) where point-based analysis is less pertinent.…”

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confidence: 99%