Live-Cell STED

 

Recording living cells is a challenge all by itself. Additionally, when doing live-cell superresolution imaging on living cells, there is a multitude of things to take care of at the same time.  

To enable you to focus on getting the best images, Abberior Instruments offers a unique live-cell package with its STED and RESOLFT microscopes:

All components are fully integrated into our software Imspector.

The full package is in its entirety only available from Abberior Instruments.

Imaging of a Region-of-Interest (ROI) Queue

Below is an example where the Abberior Instruments Live-Cell Package has been used to great effect to consecutively record five pre-defined ROIs. A formidable 100 successive frames where recorded for each region in 50-second intervals before moving on to the next ROI. Thus, each ROI was under the microscope for approx. 1.5 hours and the total acquisition time was about 7.5 hours. During this time our easy3D STED microscope ran completely autonomous.

This would not have been possible without all the contents of the Live-Cell Package: Pulsed lasers and RESCue reduced bleaching, the autofocus worked to keep the focus stable, we used water objective lenses and heated the sample to 37°C and Overnight Control assumed responsibility for automatization.

Five regions of interest were selected in a living sample stained with SiR-Tubulin.

Results

These are the results condensed into five videos from the five ROIs. Crisp STED images at physiological conditions over a time course of more than seven hours.

Live-cell Confocal and Live-cell STED images of SiR tubulin labelled mammalian cells; ROI 1, In total ~1.5h (100 frames)

Live-cell Confocal and Live-cell STED images of SiR tubulin labelled mammalian cells; ROI 2, In total ~1.5h (100 frames)

Live-cell Confocal and Live-cell STED images of SiR tubulin labelled mammalian cells; ROI 3, In total ~1.5h (100 frames)

Live-cell Confocal and Live-cell STED images of SiR tubulin labelled mammalian cells; ROI 4, In total ~1.5h (100 frames)

Live-cell Confocal and Live-cell STED images of SiR tubulin labelled mammalian cells; ROI 5, In total ~1.5h (100 frames)

Further Information