As such, live-cell imaging has proven to be a crucial part of cellular analysis in fields like molecular biology, neuroscience, diagnostics, and immuno-oncology. When using cell imaging systems, researchers are able to:
But, cellular experimentation is only as powerful as your cell imager. The capabilities and configuration of the imager, multi-mode reader (conventional microplate reading combined with widefield microscopy), or microscope can greatly impact your work. With the proper cell imaging system, you can be sure:
That is to say, all of these factors will determine the success and reproducibility of imaging experiments and downstream assays.
Understanding cellular behavior, response, and structures of both single cells and populations is made possible with the advancement of cell imaging techniques and time-lapse imaging. Compared with fixed cell microscopy, live-cell imaging greatly increases the information available to life science researchers on how stimuli affect the life and death of cells.
Cell imaging systems and methods are suitable for a range of cell-based assays and applications, including:
As mentioned, cell analysis and imaging applications can range from basic cell counts and confluence for culturing or assay preparation to automated assays and three-dimensional culture experiments, aiding many different types of research. Depending on the application needs of your lab or facility, there are several factors to consider.
It’s important to consider what type of microscope you’ll need when performing cellular imaging. Imagers come equipped with a variety of microscopes, it just depends on what type of experimentation you’re carrying out.
From simple imaging technology, like brightfield microscopy, to multi-channel fluorescence imaging, you have a wide range of options. This includes phase contrast, confocal, color-brightfield, and quantitative phase-contrast microscopy.
It’s also important to consider whether you need automated imaging capabilities for your research and assays. Cell imaging systems, depending on the type of microscope equipped, can provide automation that increases throughput and screening times.
Another factor to consider is visualization. Specifically, the live and fixed cell imaging capabilities of the system. In some cases, it’s only necessary to have an imager capable of analyzing fixed cells, but these days many labs and facilities work with live cells since they can extract more real-time data from an active sample.
For end-point measurements on cells or fixed tissue, a benchtop imager with fluorescent capabilities is usually sufficient. Yet, when working with live cells, an imager that can be integrated with an incubator is ideal.
The incubator moderates a number of important environmental factors that aid in the success of your experimentation by controlling evaporation:
A number of cell imagers can be placed directly in a dedicated incubator, while other systems have built-in incubation capabilities. While a dedicated incubator can provide the most stable conditions for live cells, it takes up additional lab space and typically generates higher costs.
Comparatively, built-in controls not only protect the cells, they allow you to record the environmental conditions present for any cross-checking purposes in case the experiment produces abnormal results.
Additionally, an injection port for reagent and drugs for immediate kinetic analysis is often important to these longer, live-cell experiments.
The optical resolution of a cell imaging system is important no matter what. Depending on the application being performed, you should consider each system’s image resolution, which is set by the objective, or objective lens, of the imager’s microscope. It is located closest to the object and relays a real-time image of the object to the eyepiece.
When performing cell counting and confluence, an objective of 4x or 10x typically delivers the best resolution. However, if you’re performing applications that require high resolution, such as intracellular assays, you’ll need to make sure the imager has the required optical resolution.
If you’re working in a lab or facility that performs high-throughput or high-content experiments using microwell plates, or you wish to expand your research capabilities, having a cell imager that is compatible with a microplate or other automation can greatly benefit you.
Being able to observe and analyze all cells within a well is incredibly important for a number of reasons. It aids in effective cell counting and confluence and offers a huge benefit to any cytometry applications, including viability and cell death measurements.
A cell imager with whole-well imaging capabilities will reduce the amount of images per well and speed up image tiling and stitching.
Cell count is incredibly important to the cell imaging process. Accurately quantifying cells helps monitor cell health and proliferation rate and aids in cell-based assay preparation. Being able to reliably and consistently count cells is also critical when studying how modulators or drugs affect cell health.
While cell counting is a fundamental part of all cell imagers, it can be highly valuable to invest in an imager that can perform confluence analysis, as this is a powerful measurement of cell viability and health.
Cell confluence, the percentage of a culture dish surface covered by adherent cells, is an exceptional alternative to label-free cell counting and is a useful tool for measuring cytotoxic effects.
Furthermore, knowing the confluence level can be an important and routine step in your workflow that also ensures overall experimental reproducibility.
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