eBook: Advances in Cancer Immunology Research - 51

es are automatically captured in brightfield and the
area is quantified using an advanced texture algorithm.
Areas of pixel variation are identified by the
system as a spheroid in the image compared to the
uniform pixel intensity of the background area. This
analysis can be utilized to assess spheroid growth
and quantify the invasion area over several days in
response to drug treatments.
Celigo performs rapid simultaneous imaging and
analysis, capturing and quantifying spheroid images
in every well of a 96- or 384-well plate in 5-10 minutes,
eliminating the need for additional image analysis
software.
Fluorescent microscopy providing
scalable in-depth data
Additional tools can also be used to access spheroid
health and gain multi-parametric information.
Spheroid viability can be quickly assessed by the
use of membrane-impermeable, nucleic acid intercalating
dyes, such as propidium iodide. These
dyes are only able to enter cells with compromised
membranes, providing information regarding the
number of dead cells in culture (Figure 4a, Shen,
et al, 2017).
Figure 4. Propidium Iodide has been used to determine spheroid health after various treatments (Figure 4a, Shen, et al)
Calcein AM dye with green/red fluorescence was used for a rapid overall health assessment readout for each spheroid (Figure
4b, Kessel, et al). A Caspase 3/7 reagent provides further information on cell death mechanisms in response to various drugs
(Figure 4c, Kessel, et al, 2020).
51

eBook: Advances in Cancer Immunology Research

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