Rapid strides in life sciences technology in the past few years have made it possible to utilize live cell imaging for a variety of applications, from cancer research to drug development. Live cell imaging technology allows the study of cellular processes at the molecular level over a period of time.

While there are many applications and microscopy methods available for live cell imaging, researchers often struggle to maintain sample integrity while they are acquiring images with sufficient resolutionduring the experiment. These 2 factors are critical for live cell imaging experiments.

Live Cell Imaging Experiment Tips to obtain relevant and reproducible data

Here are a fewbest practices to maintain the integrity of your sample and run a successful live cell imaging experiment.

Image AcquisitionUse robust autofocus modes.Minimize the power of the illuminating light source and exposure time.Use image deconvolution algorithms while using widefield microscopy systems.Use confocal imaging systems that allow high-power laser light sources and binning to be used.Employ powerful analysis software to get data that is reproducible and meaningful.

Sample preparation - Use the correct media formulation

While you make sure that media formulations have the appropriate nutrients for the growth and development of cells, do not forget to keep an eye on pH, buffering capacity, and osmolarity if you want to run a successful live cell imaging experiment.

Changes in pH and osmolarity can affect cell expression, phenotype, and how they appear and behave. Optimal environmental conditions can help you avoid unwanted changes to the media.

A lot of factors within media can cause auto fluorescence or high background signal in the images. Use phenol red-free media and reduce serum concentration to reduce such phenomena.

Sample imaging - Control and maintain proper environmental conditions

Maintain the right environmental conditions, like temperature, humidity, and gas, for the samples throughout the live cell imaging experiment.

Temperature variations can cause expansion and contraction of the microplate or other culture vessels and hence, focus drift. A stable temperature means cellular health as well as less risk of losing focus over time. You can also reduce the risk of focus drift by ensuring the microplate is seated on the instrument stage to allow for thermal equilibration before configuring acquisition settings. If you need to add media to wells in the time-lapse experiment, it should be the same temperature as the media in the imaging vessel.

Maintain proper humidity to prevent media evaporation and osmolarity changes. Both can negatively affect the cells and your assay. Use a hypotonic solution if humidity control is not possible.

Control CO2 levels to regulate sample pH.  If this is not possible, you can use synthetic buffers, such as HEPES, to maintain pH but don’t forget to verify the compatibility between the cells and the synthetic buffer before use.

Use Phase Holographic Imaging

Phase Holographic Imaging, PHI Holomonitor® is a non-invasivelive cell imaging and analysis tool. It is a compact microscope that can continuously image, monitor and, quantitatively analyze both single cells and populations inside a normal without any labels or stains.

Its small foot-print, and ease of use, has made the HoloMonitor® M4 a staple in labs working with cell cultures for obtaining real-time, time-lapse quantitative data as well as 3D images and/or movies of your cell samples.

To learn more about live cell imaging, visitNexus Scientific, authorized distributors of PHI Holomonitor® in the USor call857-217-0936.