ZEISS LSM 900 with Airyscan 2

To minimize photobleaching and damage to a live sample, it is useful to reduce acquisition time and to use minimal laser power. LSM Plus helps to improve the signal-to-noise ratio as well as the resolution of structures such as spindle fibers.

In this example, 100 Z-stacks were acquired with LSM 900 on Celldiscoverer 7 over 29 minutes. The images show a maximum intensity projection of 38 Z-planes. Cells expressing H2B-mCherry (red) and α-Tubulin-mEGFP (cyan).

LSM Plus helps to improve signal-to-noise ratio when imaging large volumes to be rendered in 3D. Visualization of the vasculature (green) and red blood cells (magenta) by transgenic reporter expression, lateral view, anterior to the left.

A 300 µm Z-stack with 81 planes over three tiles was imaged with LSM Plus applied.  The tiles were stitched and rendered in 3D with ZEN – powered by arvis^®.

Sample courtesy of B. Schmid, DZNE Munich, Germany

Images were acquired with LSM 900 on ZEISS Celldiscoverer 7 using confocal GaAsP detectors (top row) and Airyscan 2 in HS mode (bottom row). Confocal images with LSM Plus (top, right) enhancing SNR and improving resolution of mitochondrial structures. Airyscan Joint Deconvolution (bottom, right) resolves the inner and outer membrane architecture even better compared to Airyscan HS (bottom left). Stained for mitochondrial outer membrane protein Tom20 (Green, Alexa Fluor-488) and mitochondrial inner membrane protein ATP5a (Magenta, Alexa Fluor-647).

Lateral line primordium migration and deposition of immature neuromasts in a Zebrafish embryo (Danio rerio).  Animals were anesthetized and embedded using low concentrated agarose in a glass bottom petridish. Initial camera based imaging allowed for a quick and easy sample navigation (top) combining Phase Gradient Contrast with fluorescence acquisition. Subsequent high resolution imaging with Airyscan 2 in Multiplex mode was done on individual positions identified in the widefield image (white boxes). The gentle and fast image acquisition that is inherent to the Airyscan 2 Multiplex mode is very beneficial for this kind of application. The animal is unperturbed by the imaging while images with a very high signal to noise ratio as well as level of detail can be acquired at the same time.

Human lung epithelial cell line A549 stained with Mitotracker Orange (mitochondria) and SIR-DNA (nuclei).

The acquisition seemlessly combines two imaging modes – the fluorescent channels were captured in confocal mode using highly sensitive GaAsP detectors while the Phase Gradient Contrast is camera based.

A timelapse of 2.5 h was acquired using a 40× magnification with a numerical aperture of 0.95.