Super-resolution microscopy enables the visualization of cellular components at resolutions down to 10 nm. However, a critical bottleneck is that the movies last only for seconds, and miss dynamics that are essential to truly understand how these structures function and interact in health and disease. Recently, the Toomre/Schepartz lab have developed a set of high-density, environment-sensitive (HIDE) membrane probes that enable long time-lapse, live-cell nanoscopy of discrete cellular structures and organelles with high spatiotemporal resolution. However, such HIDE probes could only visualize single target. Here, we described the development of two-color HIDE probes for long-time lapse Stimulated emission depletion (<i>STED</i>) imaging. HIDE probes targeting the ER, mitochondria, and/or plasma membrane facilitated two-color STED imaging of organelle dynamics for up to 8 minutes, far exceeding the movie length of previous examples. We further applied the new method to image primary and hard-to-transfect cell lines to prove the general utility of this method.

Two-color long time-lapse super-resolution imaging with lipid probes

2019 Spring National Meeting

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