A novel miniature cell retainer for correlative high-content analysis of individual untethered non-adherent cells

Abstract

The importance of research involving non-adherent cell lines, primary cells and blood cells is generally undisputed. However, the task of investigating the complexity and heterogeneity of these cells calls for their long-run monitoring at a single-cell resolution. Such a capability is currently unavailable without having to use disruptive cell tethering. The present Cell Retainer (CR) concept enables high-content correlative multi-parametric measurements, from the functional to molecular level, of the same living individual non-adherent cells within a population. Thereby, despite extensive long-term bio-manipulations, the cells preserve their identity without tethering. Several exemplary experiments, using a microscope-slide-based version of the CR, are presented, which could not be performed by other state of the art methods.

Identification of antigen‐specific B cells by concurrent monitoring of intracellular Ca2+ mobilization and antigen binding with microwell array chip system equipped with a CCD imager

“…Cytomics is becoming increasingly important for analyzing cell conditions and cellular responses to stimuli from extracellular milieu, which will contribute to drug discovery as well as predictive medicine. Cell microarray-based analysis offers one way for analyzing cytomics; it has been conventionally performed with a single parameter (11)(12)(13)(14)(15)(16)(17)(18). Here, for the first time, we present a method of two-parameter cell microarray-based time-lapse analysis.…”

“…After that, the MAC-CCD system detected the position of each microwell on the chip again. Fifteen seconds after the stimulation, the fluorescence images of the fluo-4 signals were acquired on the whole microwell area every 3 s for an additional 45 s (scanning points [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20]. The cells inside the wells did not move out of the wells after four gentle washes.…”

“…To date, some cell microarray technologies using microwells have been reported for analyzing a large number of nonadherent cells (11)(12)(13)(14)(15)(16). These microwells are designed to capture single cells, enabling the tracking and analyzing of cellular functions at the single-cell level.…”

Single lymphocyte analysis with a microwell array chip

“…Recently, Deutsch et al reported a live cell array to perform cell-based assays on thousands of individual cells (7). Microcavities were arrayed in a honeycomb-like structure and could monitor cell responses, such as reactive oxygen species generation, on a single-cell basis (23).…”

Technology Advancement for Integrative Stem Cell Analyses

“…1a). 66,[71][72][73] These systems use computer-controlled, pneumatically driven systems to manipulate small quantities of fluids precisely through multiple modules. The utility of these systems for the detection and amplification of DNA or RNA from individual cells has been described.…”