Asymmetric cell division in a live zebrafish embryo. Credit: Paula Alexandre via Wellcome Collection
by Michelle Taylor, Editor-in-Chief
Live cell imaging is an important analytical tool in many laboratories. The ability to visualize protein dynamics in vesicles, organelles, cells, and tissues has provided new insights into how cells function in healthy and disease states. These insights include the spatiotemporal dynamics of processes like mitosis, embryonic development, and cytoskeleton changes.
But, biological imaging of sensitive samples is complex. The microscopy method utilized should minimize photobleaching, phototoxicity and other potentially harmful outcomes that can irrevocably damage a sample. At the same time, the technique must deliver ultra-sharp resolution and allow researchers to peer deep into specimens.
Tip No. 1: Consider Sensitivity, Speed and Sample
When choosing an optical microscopy system, there are three important variables: sensitivity of the detector (signal-to-noise), the speed required for image acquisition, and type of specimen being analyzed.
To optimize signal-to-noise, the combination of filters selected for imaging of live cells should closely match the spectral profiles of the fluorophores used for experiments. Signal-to-background ratio can be optimized by using reagents that reduce extracellular fluorescence and increase fluorophore photostability.1 It is important to image in media that have been specifically designed for maintaining cell health while reducing or eliminating background fluorescence. The addition of a background suppressor compatible with live cells can also help reduce extracellular background fluorescence and eliminate the need for a wash step. Antifade mounting media for live cells can be applied to samples to reduce photobleaching of fluorophores, preventing signal loss with multiple or long exposures.1
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