Page 12 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

The IncuCyte system is compatible with multiple different types • Documentation of cell morphology
of cell culture vessels and can monitor multiple vessels at a time. Changes in cell morphology due to cell seeding density
Data consists of objective, image-based growth metrics to capture or phenotypic drift can be identified using a range of
transient and time-dependent events. magnifications to capture fine details of cells and spatial
coverage of cell populations.
In order to establish quality cell cultures and improve • Accurate assessment of overall monolayer
experimental outcomes in downstream assays, various aspects By imaging multiple areas of your vessel or using a whole-well
of cell culture need to be tightly controlled, optimized and imaging mode, spatial variations in cell distribution can be
documented. quantified and then reduced.

• Cell seeding densities
• Growth conditions To improve assay quality and consistency of kinetic assays, it
Prior to utilizing cells in quantitative assays, it is important is important to minimize variations in cell seeding densities
to optimize and define cell culture regimens. Documenting due to pipetting errors during plate seeding which result in
variations in cell growth due to factors such as; lot-to-lot differential growth rates. Measuring confluence before utilizing
differences in cell culture media, changes in media component assay plates eliminates assay variation, resulting in reproducible
concentrations over time due to degradation, and inconsistent conditions.
feeding schedules can all be identified using label-free phase
image segmentation techniques that generate confluence • Documenting time-dependent variables
measurements. Assessment of cell behavior and growth in association with cell
treatment times can define experimental conditions to ensure
consistent results.

Cell Culture QC Assays
Measuring proliferation to optimize growth conditions and seeding densities

The IncuCyte system can be used to track cell proliferation over time using label-free
phase segmentation (confluence) metrics (Figure 2), allowing for the assessment of cell
growth as well as monitoring cell culture and assay optimization.

A B

C

Figure 2. T-flask vessel view shows
location of images acquired (A).
Proliferation time-course for HT-1080
human fibrosarcoma cells is shown
(B). (C) HD-phase time-lapse images
of HT-1080 cells (confluence mask
overlaid; right panels).

10