Page 86 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

Key advantages:
• Quantify label-free growth and investigate morphology of • Generate reproducible data suitable for pharmacological analysis
multiple spheroids grown on ECM inside a standard tissue with lab-tested and cost-conscious protocols, high quality
culture incubator, ensuring improved physiological relevance. images, and unbiased analysis.

• Multiplex kinetic viability and toxicity measurements using • Rapidly create concentration response curves from thousands of
IncuCyte® non-perturbing Cell Health Reagents to investigate images generated in 96-well plates.
drug mechanisms of action.

Application of IncuCyte® S3 Multi-Spheroid live-cell assays

Combining the IncuCyte® DF Brightfield image acquisition with a the incubator while images are taken, it is possible to visualize
3D multi-spheroid cell model on ECM allows for monitoring and and quantify spheroid growth, measure cell health, and better
quantification of changes over time and under more physiological understand mechanisms of drug action.
relevant conditions. As the spheroids remain undisturbed within
Label-free image acquisition using novel DF-Brightfield reveals morphology and enables
quantification of spheroid growth over time

Introduction of a new, enhanced depth of focus brightfield The novel DF Brightfield image capture provides high-quality
(DF Brightfield) image acquisition enables imaging of multiple images of multi-spheroids formed from a range of tumor cells
tumor spheroids on a layer of extracellular matrix (Figure 2). This (Figure 3). Within just three days, different tumor cell lines begin
enhanced image acquisition results in high contrast brightfield to show distinctive morphology, demonstrating the physiological
images that can be reliably segmented and analyzed using relevance of the models.
IncuCyte’s purpose-built processing algorithms.

Traditional Brightfield DF Brightfield

Figure 2. IncuCyte’s proprietary image acquisition technique, DF Brightfield for 3D Cultures, generates high contrast, extended depth of focus images when
used in conjunction with the IncuCyte S3 Multi-Tumor Spheroid protocol. Subsequent segmentation and analysis with IncuCyte S3’s image processing
algorithms results in reproducible data without operator bias.

Lung A549 Ovarian SKOV-3 Breast MCF-7 Breast MDA-MB-231
"Round" "Round" "Round" "Stellate"

HD Phase
Image

DF-BF
Image

Figure 3. High quality HD Phase and DF Brightfield (DF BF) images of multi-spheroids (MS) formed from a range of tumor cell lines (5 d post seeding) on
a Matrigel® base. Three day post seeding, A549, SKOV-3 and MCF-7 cells formed round aggregates, while MDA-MB-231 MS exhibited stellate branching
distinctive of an invasive morphology.

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