Cell chip array for microfluidic proteomics enabling rapid in situ assessment of intracellular protein phosphorylation

Abstract

We discuss the ability to perform fluorescent immunocytochemistry, following cell fixation, using a microfluidic array of primary, nonadherent, single CD34+ stem cells. The technique requires small cell samples and proceeds with no cell loss, making it well-suited to monitoring these rare patient-derived cells. The chip allows us to correlate live cell dynamics across arrays of individual cells with posttranslational modifications of intracellular proteins, following their exposure to drug treatments. Results also show that due to the microfluidic environment, the time scale of cell fixation was significantly reduced compared to conventional methods, leading to greater confidence in the status of the protein modifications studied.

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

“…Faley et al [29] described a microfluidic technique by which rare chronic myeloid leukemia CD34+ hematopoietic stem/progenitor cells could be individually immobilized, followed through live imaging and assessed for protein post-translational modification, and rapidly fixed in situ (Fig. 1A).…”

Single-Cell Omics Analyses Enabled by Microchip Technologies

“…Finally, cell immunostaining is a straightforward technique for intracellular protein detection and can be readily integrated in microfluidic chips. Single cells can be isolated using a hydrodynamic trapping array, electroactive microwell array, or digital microfluidic device and stained by immunocytochemical reagents (161)(162)(163)(164). the phosphorylation states of platelet-derived growth factor receptors as well as the downstream signaling protein Akt to investigate the effects of chemical stimulation.…”

Quantitative profiling of integrin αvβ3 on single cells with quantum dot labeling to reveal the phenotypic heterogeneity of glioblastoma

“…The development of precise analytical techniques has greatly expedited the study of genomics, 1,2 transcriptomics 3-5 and proteomics, [6][7][8] as well as epigenomics 9 at the single-cell level. A large number of studies have revealed diversity and heterogeneity within the same type of cells, thus providing invaluable insights into the complexity of biological processes and disease mechanisms.…”