Striking a Balance for Perfect Images
Advanced microscope systems enable researchers to perform high-resolution live-cell imaging, while maintaining cellular health.
Scientists employ live-cell imaging to capture dynamic cellular processes over time. While they can observe unlabeled cells, researchers commonly label cell structures or biomarkers with fluorophores to provide subcellular information about their samples. However, live-cell imaging presents them with several challenges, making it a complicated technique to master.
Researchers must balance acquiring high-quality images with ensuring that the cells remain healthy. Optimal imaging settings, such as high illumination intensity, long exposure times, and short intervals between images, are often detrimental to cellular health.1 Conversely, the gentle settings required for ideal cellular health frequently produce image series with inadequate spatial and temporal resolution.
Further exacerbating this problem, cellular health is also affected by environmental conditions, including temperature, pH, and nutrient or ion concentrations.2 Temperature fluctuations also cause drift, where a microscope loses its focus on a chosen focal plane, which often derails experiments.
Additionally, most microscopes acquire a separate image for each fluorescence channel. If the cells and internal structures move during this process, the images appear flawed when the software overlays the channels, which hinders spatial-temporal correlation analysis between the fluorophores.
Microscope systems, such as Mica from Leica Microsystems, overcome these challenges through innovative design and intuitive controls. When equipped with the live-cell imaging package, this automated system stabilizes the temperature, carbon dioxide levels, and humidity of the imaging environment. In addition to its drift correction and widefield and confocal modes, the instrument acquires four fluorescence channels simultaneously, producing perfectly merged images. Microscope systems like this one improve live-cell imaging by balancing cellular health and image quality.
- Jensen EC. Anat Rec. 2013;296(1):1-8.
- Ettinger A, Wittmann T. In: Methods in Cell Biology. Vol 123. Academic Press; 2014:77-94.