Live cell imaging for phenotypic assessment

As opposed to standard endpoint methodologies, live cell imaging & analysis platform allows for visualization and quantification of cell behavior overtime. This time-lapse and fully kinetic monitoring up to several days on living cells provides thus real-time insight into a variety of active biological processes and enables quantitative data analysis. Thanks to these features, live cell imaging allows to assess a variety of live-cell assays in a faster and automated way. In assessing compounds effects, the platform allows thus characterization of cell behavior under different treatment conditions in dose- and time-dependent manner.
 

Key features and advantages of live cell imaging

  • The well-suited platform for short- and long-term kinetic live cell studies: to characterize cell behavior and get a complete time- and dose-dependent pharmacological profiling.
  • Compatible with phase-contrast label-free and 2 fluorescent phenotypic assessments.
  • Powerful image visualization & time-course analysis, enabling real-time decision making.
  • Able to multiplex with additional functional measures e.g. cytokines, or endpoint immunofluorescence detection & analysis.
  

Platform

IncuCyte® S2 Live-Cell Analysis System

  • Phase-contrast & fluorescent time-lapse monitoring

Cell models

  • Live cell cultures (primary & cell lines)
  • Live cell co-cultures (primary & cell lines)
  • Many culture vessels, in addition to 96-w & 384-w plates (up to 6 at a time)

 


Illustrative data

Immune cell-mediated killing of H1299 lung cancer cells
H1299 tumor cells stably expressing a nuclear red fluorescent probe are cultured with inactivated or aCD3-activated hPBMC.

 

A. aCD3 induces hPBMC-mediated apoptotic killing of target tumor cells. Live cell real time detection images of apoptotic H1299 tumor cells at 72h post-culture with inactivated and aCD3-activated hPBMC. Tumor cell count (nuclear red probe-expressing cells, (a, b, d, e)) and apoptosis (caspase 3/7 green fluorescent probe, (a, b, d, e)) are monitored, and segmentation masks & image analysis are performed (c, f) to quantify tumor cell apoptosis (i.e. caspase 3/7 green fluorescent probe within the nuclear red probe-expressing cells – yellow objects). In e arrows indicate apoptotic tumor cells and arrowheads apoptotic non-tumor cells.

B. Kinetic and dose-dependent effect of aCD3 in inducing hPBMC-mediated apoptotic killing of H1299 cancer cells. Tumor cell apoptosis is quantified and results normalized to tumor cell count and plotted over the time.

H1299 tumor cells stably expressing a nuclear red fluorescent probe are cultured with inactivated ...

Dose- & time-dependent T Jurkat cell migration to SDF1 Alpha
H1299 tumor cells stably expressing a nuclear red fluorescent probe are cultured with inactivated or aCD3-activated hPBMC.

 

A. aCD3 induces hPBMC-mediated apoptotic killing of target tumor cells. Live cell real time detection images of apoptotic H1299 tumor cells at 72h post-culture with inactivated and aCD3-activated hPBMC. Tumor cell count (nuclear red probe-expressing cells, (a, b, d, e)) and apoptosis (caspase 3/7 green fluorescent probe, (a, b, d, e)) are monitored, and segmentation masks & image analysis are performed (c, f) to quantify tumor cell apoptosis (i.e. caspase 3/7 green fluorescent probe within the nuclear red probe-expressing cells – yellow objects). In e arrows indicate apoptotic tumor cells and arrowheads apoptotic non-tumor cells.

B. Kinetic and dose-dependent effect of aCD3 in inducing hPBMC-mediated apoptotic killing of H1299 cancer cells. Tumor cell apoptosis is quantified and results normalized to tumor cell count and plotted over the time.

Chemotaxis monitoring & quantification by live cell imaging. T Jurkat cells migrate towards ...

Neurite outgrowth monitoring in mature cortical neurons
Glutamate toxicity on neurite outgrowth of mature rat cortical neurons by live cell imaging. 10 DIV-old cortical cultures are exposed to increasing glutamate concentrations, and neurite outgrowth is kinetically monitored over 96h by live cell imaging. Glutamate dose-dependently affects the neuritic network over the time.

Glutamate toxicity on neurite outgrowth of mature rat cortical neurons by live cell imaging. ...