This C. elegans line was created by C. W. Okafornta, Dresden University of Technology, Germany. It is equipped with fluorescent marker proteins for cell membranes (PH::mKate2), nuclei (H2B::mCherry) and centrosomes (gammaTub::GFP). This allows for visualization of cellular structures to analyze cell division cycles and cell compartmentalization during live embryonic development.
Image data for this use case was captured with ZEISS Lattice Lightsheet 7. This system is designed for gentle, volumetric live cell imaging, making it the ideal instrument for observing delicate structures over long periods of time. The C. elegans embryos were imaged using the green channel (centrosomes) and red channel (cell membranes and nuclei). Live acquisition was performed for almost 2 hours with one volume every 30 sec, 476 planes per volume for a total of 120,000 images.
When using lattice light sheet microscopy, the sample is imaged at an angle, due to the geometry inherent to this technology. Consequently, the raw data must be processed (deconvolved and deskewed) in ZEN (Figure 2) before further analyses can be performed.
The processed .czi file from ZEN was directly imported into ZEISS arivis Pro (formerly Vision4D) and converted into the arivis .sis file format, a format specifically designed for fast handling of large data sets. We then reduced the data size by using only every second time point, resampling the data set to 50% in z dimensions and converted to data set to 8-bit. The final data set has 2 channels, 79 slices, 60 time points, and a size of ~0.9 GB.