Autoimmersion Module for ZEISS Axio Observer 7​

HEK KO PEX5 cells expressing eGFP with a photocaged peroxisomal targeting signal type 1 were reconstituted with the peroxisomal import receptor PEX5. A light induced conformational change of the photocage leads to exposure of the peroxisomal targeting signal. If the WT PEX5 is expressed, accumulation of the eGFP signal in the dotted peroxisomes can be monitored (top row). In case of the mutated PEX5 (bottom row), even after 18 hours, no peroxisomal import could be detected.

When working with living samples, you might not know where your event of interest may occur. To capture the uptake of nanoparticles by macrophages, many locations from a multi-well plate are acquired as well as multiple Z-planes over several hours at 37°C using re-immersion. The region shown above is a subset of the much larger dataset that was captured using automated imaging and shows the uptake of nanoparticles inside the cells (top row). The surface of the cells were also imaged to verify that the nanoparticles are inside the cells and not simple sitting on the cell surface (bottom row).