Application Note: Kinetic Quantification of Cell Proliferation Using Live-Cell Analysis
Cell proliferation assays are a cornerstone of cancer therapeutic, developmental biology, and drug safety research. Analysis of the sustained signaling pathways that underlie the progression of tumors, for example, accounts for > 12,000 manuscripts in PubMed—the majority of which use cell proliferation analysis to evaluate tumor cell growth. However, there are several technical challenges associated with this seemingly simple activity. Namely, cell growth takes time, which makes temporal monitoring and non-perturbing measurements important for high-quality results.
Live-cell analysis is well suited to address these challenges by providing valuable information without interfering with cell growth, thus ensuring reliable, reproducible, and comparable data. Live-cell analysis enables the real-time quantification of living cells’ behavior through time-lapse imaging, integrated image analysis, and on-the-fly data visualization for cell-based experiments over days, weeks, or even months. The need for multiple assays with different end points is therefore eliminated, allowing you to continuously collect data without the risk of missing a key biological event. Additionally, the noninvasive nature of this technology makes it complementary to flow cytometry and multimode microplate readers. It can also be used in combination with label-free or specialized labeling approaches to expand into more complex assays.
Multiple strategies exist for kinetic measurement of proliferation using the Incucyte® Live-Cell Analysis System. Utilizing label-free or fluorescent assays depends on the specific scientific question being asked and cell models studied. Continuous live-cell assays for both adherent and non-adherent cells is possible because cells remain stationary inside a standard tissue culture incubator while the Incucyte® optics move. Incucyte® Live-Cell Imaging and Analysis enables noninvasive, label-free measurement of cell growth based on area (confluence) or cell number (count) metrics, both of which are generated via segmentation (masking) of high-quality phase images. For low-contrast cells that can be difficult to identify and quantify in phase contrast images, Incucyte® Nuclight Reagents can be used to fluorescently label nuclei. Fluorescent Incucyte® images can then be acquired over time and analyzed to generate nuclear counts and derive doubling times in either mono- or co-cultures.
Combining label-free image analysis with the Incucyte® Live-Cell Analysis System, non-perturbing fluorescent reagents, and easy-to-use software tools provides a powerful solution for kinetic cell proliferation measurements and pharmacology assays.
In this application note, we explore:
- Kinetic, label-free analysis of cell proliferation and cell counting
- Continuous live-cell proliferation for non-adherent cells
- Fluorescence labeling to track proliferation
- High throughput compound testing capabilities
