What To Consider When Choosing The Right Microscope For Live Cell Imaging System

To perform live-cell imaging experiments successfully, using the right approach is critical. When choosing a suitable microscope for your live‐cell imaging, the following aspects should be considered: Specimen viability, image acquisition speed (temporal resolution) and required resolution in all three dimensions.

During live cell imaging, certain environmental conditions must be maintained to avoid detrimental physiological changes. In order to capture physiologically relevant cellular dynamics, live cell experiments require specific environmental conditions, including temperature, pH (via CO2), and humidity control. Furthermore, some experiments may even require hypoxic conditions. Modern incubation systems not only tightly control environmental conditions, they can also provide detailed data reports and alert users to temperature, gas, or humidity variations during the course of an imaging experiment. To minimize or avoid the effects of photodamage, getting the right balance between sensitive detection, accurate label separation (if using >1 label) and the lowest light dosage for excitation is crucial.

For live cell experiments, high speed acquisition is often critical, in particular for the study of fast dynamic processes such as vesicle observation. Using optical filters results in speed limitations due to the necessity for sequential imaging when changing filter sets for each color, used to study the interaction of multiple components. Gathering images sequentially requires more time than simultaneous image gathering and, as a result, rapid specimen motions can be missed during acquisition, as each color has a longer time interval from one image to the next. On top when the direct comparison between two or more colors is of essence, the signals may have moved even between the individual acquisition of the fluorophores, complicating the interpretation of the data.

Multiple technologies are available for acquiring images in 3 dimensions over time. The choice of system depends on your experiment and whether higher speed or less sample illumination during imaging is your priority when acquiring the desired 3D resolution. Choosing the most appropriate system has traditionally required you to make a choice between a camera-based or confocal live cell imaging system, however modern solutions can provide both modalities in an integrated way.