Brightfield and Fluorescent Image Analysis for Screening Applications using the Celigo Adherent Cell Cytometer

Olivier Dery, Sarah Kessel, Kristi A. Hohenstein Elliott, Cory Peterson, and Fredrik Kamme

The Celigo adherent cell cytometer is a multi-channel imager
well suited for all image-based screening applications. The
flexibility and ease-of-use of the Celigo facilitates the
optimization of cell-based assays and the subsequent
evaluation of assay performance. Using large field-of-view
optics and a proprietary high speed imaging system, the Celigo
rapidly images multiple fields-of-view without moving the plate.
Nearly all microplate formats (from 1536-well to 6-well plates)
can be fully- or partially-imaged in brightfield and 3 fluorescent
channels. Additionally, unique brightfield optics allow accurate
segmentation of cells right to the edge of the well and a rich flow
cytometry-like gating interface accommodates a wide variety of
cell-based assays. We present here whole-well imaging and
label-free brightfield cell counting applications that allows the
Celigo user to generate growth curves over time and monitor
cell counts and confluence at the individual well level. These
brightfield applications permit the screening of drugs altering cell
proliferation and growth rate. We also highlight the capacity of
the Celigo to image large objects making it suitable for analysis
of large structures such as embryoid bodies, tumor spheres or
small organisms. Finally, the analysis of complex samples by
imaging is a challenge for high-content screening applications.
The Celigo software provides multi-channel analysis of
brightfield and fluorescent images and means of identification of
cells based on intensity and morphology parameters. Moreover,
a powerful yet easy-to-use gating interface allows the analysis
of cell populations similarly to flow cytometry. Data also shows a
variety of fluorescent assays for assessment of cell health, DNA
synthesis, cell cycle or expression of cell surface markers.
Overall the brightfield and fluorescent imaging capability of the
Celigo combined with powerful analysis features and a potential
integration with automation make it suitable for high-throughput
screening applications.