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How to Perform Dynamic Multicolor Time-Lapse Imaging

Simultaneous imaging for absolute spatiotemporal correlation

Living HeLa cells stained with WGA-488 (yellow), SPY-Actin (cyan), and SiR-Tubulin (magenta). Instant Computational Clearing (ICC) was applied. Living_HeLa_cells_Instant_Computational_Clearing.jpg

In this article, we provide examples about the ability of Mica to do dynamic live-cell imaging. Live-cell imaging sheds light on diverse cellular events. As many of these events have fast dynamics, the microscope imaging system must be fast enough to record every detail. One major advantage of such an imaging system would be the ability to capture several fluorescence imaging channels simultaneously for their exact spatiotemporal correlation. Mica enables users to acquire the signal of up to four fluorophores at a time, either in confocal or widefield mode. With it, cellular components can be imaged at the very same point in time, without spatiotemporal mismatch.

Example: Four-Color Time-Lapse Imaging – Confocal

Here is one example showing simultaneous imaging in confocal mode.

U2OS cells stained with MitoTracker™ Blue (blue), FluoView 488 Tubulin (green), WGA-Alexa555 (yellow), and SiR-Actin (cyan). Objective is HC PL APO CS2 63x/1.20 WATER. Simultaneous acquisition. Time-lapse imaging with 5-second intervals for a total duration of 2 minutes.

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Reference

Efficient Long-term Time-lapse Microscopy, Science Lab, Leica Microsystems, 2022.

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