ZEISS Lattice Lightsheet 7

Human induced pluripotent stem cells which endogenously express mEGFP-tagged lamin B1 (AICS-0013). Images generated using AICS-0013 (LMNB1-mEGFP) from the Allen Institute for Cell Science. The overnight experiment was recorded for close to 8 hours with one volume imaged every 1.5 min.

Lamin B1 is involved in disassembling and reforming the nuclear envelope during mitosis. The formation of so-called ‘nuclear invaginations’ has been reported frequently during mitotic events. However, most research so far on these unique structures has been done with fixed cells and as a result their function is largely unknown.

The data observes cells going through mitosis throughout the whole duration. The formation and dynamics of nuclear invaginations can clearly be observed in most of the cells, throughout the complete cell cycle. The gentleness of Lattice Lightsheet 7 imaging enables imaging of mitotic events over longer time periods, preventing arrest from excitation light. Combined with the fast volumetric imaging and near-isotropic resolution the ZEISS Lattice Lightsheet 7 is the perfect tool for challenging experiments like this.

T cell expressing Lifeact-GFP. Color-coded depth projection and maximum intensity projection side-by-side. The T cell was imaged constantly for over 1 hr; one volume every 2.5 secs. Sample courtesy of M. Fritzsche, University of Oxford, UK

T cell expressing Lifeact-GFP. Color-coded depth projection and maximum intensity projection side-by-side. The T cell was imaged constantly for over 1 hr; one volume every 2.5 secs. Sample courtesy of M. Fritzsche, University of Oxford, UK

Cos 7 cells transiently transfected with CalnexinmEmerald and EB3-tdTomato. EB3 labels the growing ends of microtubules and is necessary for the regulation of microtubule dynamics. Calnexin is a protein of the ER where proteins are synthesized. Cells were imaged for 1.5 hrs every 80 secs, imaged volume: 175 × 120 × 70 μm³.

Spheroids and organoids are in vitro models of organs – much smaller and simpler but easy to produce and thus for developmental biologists an invaluable tool to study organ development. Unlike cell cultures, which usually consist of a monolayer of cells only, cells in spheroids / organoids form three-dimensional structures, allowing for the investigation of cell migration and differentiation inside 3D cell models. With lattice light sheet microscopy, imaging the development and self-organization of organoids becomes reality. Here, we can see a 3D rendering of a spheroid consisting of cells expressing H2B-mCherry (cyan) and α-Tubulin-mEGFP (magenta). Not every cell is labelled.

DeltaD-YFP transgenic zebrafish embryo (Liao et al. 2016, Nature Communications). Fusion protein driven by a transgene containing the endogenous regulatory regions, expression in the tailbud and pre-somitic mesoderm. Signal visible in the cell cortex, and in puncta corresponding to trafficking vesicles (green). Nuclei in magenta. The embryo was imaged for 5 minutes constantly; one volume (150 × 50 × 90 μm3) every 8 sec.  Sample: courtesy of Prof. Andrew Oates, EPFL, Switzerland.