ZEISS Celldiscoverer 7 with LSM 900 and Airyscan 2

ZEISS Celldiscoverer 7 is an automated live cell imaging system designed to make complex microscopy simple enabling high-throughput applications, but what about the data? With easy to develop Image Analysis Pipelines in arivis Vision4D you can even tackle those samples which traditionally produce high demand on analytical techniques. Once a pipeline has been created and optimized, it is then possible to scale up the analysis by use of the server-based VisionHub platform enabling users to run batch analysis and have results readily available within the web-based user interface.

In our new series “From Image to Results”, explore how data captured with the ZEISS Celldiscoverer 7 can be combined with a powerful Image Analysis Pipeline delivered by arivis Vision4D to scale up analysis of organoid formation.

Kidney Tubule Organoids grown from mIMCD3 cells. Plan-APOCHROMAT 20x/0.7; 0.5x tube lens. Airyscan SR mode.

Celldiscoverer 7 enables confocal image acquisition of large scanning fields at high resolution. Imaging of tubular structures in multiwell plates is accomplished at unprecedented speed, allowing for high-throughput applications such as drug screening.

HeLa cells infected with Shigella flexneri expressing mCherry at 3 hours post infection to investigate cytoskeleton rearrangements during intracellular infection eg. formation of septin cages, actin tails. The sample was stained with DAPI (blue, nuclei), anti-SEPT7 antibodies (green, septin filaments), and phalloidin (magenta, F-actin). Maximum Intensity Projection acquired using the Celldiscoverer 7 using the 50x water object and the 0.5x magnification optics. Sample courtesy of A. López Jiménez, London School of Hygiene and Tropical Medicine, UK.

Zebrafish line Tg(lyz::dsRed). Observing emergency granulopoiesis in a 4 day post-fertilisation zebrafish embryo. The innate immune system induces this response to replenish the neutrophil (dsRed) population 48 hours post infection with a non-lethal dose of Shigella flexneri. Maximum Intensity Projection acquired using the Celldiscoverer 7 in widefield mode. Sample courtesy of M. Gomez, London School of Hygiene and Tropical Medicine, UK.

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.

Maximum intensity projections of the lateral line primordium tip migrating through the animal (155 z-planes).

Green: LYN-eGFP (mebranes); Red: tagRFP-T-UTRCH (actin).

Organoid from a human breast cancer cell line. The cells express GFP-labeled H2B (nucelei) and mCherry (cytoplasmic staining depicted here in grey for better visualization).

Several organoids where grown in a multiwell plate with Matrigel. Initial sample navigation was performed using the transmitted light at a low magnification of 2.5× to identify interesting organoids.

Subsequently, high resolution images were acquired using the water immersion objective with a total magnification of 50×. 61 z-planes were acquired using ZEISS Celldiscoverer 7 with LSM 900 and Airyscan 2 in Multiplex mode.

Human lung epithelial cell line A549 stained with MitoTracker™ Orange (mitochondiria) and SIR-DNA (nuclei).

The acquisition seamlessly 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.