Expansion microscopy of zebrafish for neuroscience and developmental biology studies

Abstract

SignificanceWe explore the utility of expansion microscopy (ExM) in neuroscience and developmental biology using the zebrafish model. Regarding neuroscience studies, ExM enables the tracing of cellular processes in the zebrafish brain, as well as the imaging of synapses and their biomolecular content and organization. Regarding development, ExM enables the resolving of nuclear compartments, particularly nuclear invaginations and channels, and helps relate such cellular nanostructures to proteins of the cytoskeleton during embryogenesis.

Glia Accumulate Evidence that Actions Are Futile and Suppress Unsuccessful Behavior

“…Radial astrocyte processes project ventral-laterally. Scale bar is corrected for 43 expansion (Freifeld et al, 2017;Tillberg et al, 2016) Since astrocytes support neuron health (Banker, 1980;Lyons and Talbot, 2014), we controlled for adverse effects of glial ablation on neurons. After ablations, we observed no significant difference in swim frequency, power, or velocity in freely swimming fish ( Figure 4F).…”

“…All antibody staining steps included shaking. Expansion Expansion was done as described (Tillberg et al, 2016) (see also Freifeld et al, 2017). AcX was resuspended at 10mg/mL in anhydrous DMSO, aliquoted, and stored desiccated at -20 C for up to several months.…”

Expansion microscopy: enabling single cell analysis in intact biological systems

“…After a single round of 4.5x linear expansion, Chen et al measured 1-4% distortion errors in measurements over length scales of tens to hundreds of microns ( Fig. 1E-G; [34]), an error range that has now been replicated for multiple expansion protocols applied to a diversity of other cells and tissues, including human tissues [39,[41][42][43][44]. Notably, this error varies nonlinearly with the actual measurement length; larger lengths have less relative error (Fig.…”

Expansion microscopy

“…Third, ExM needs to be tested with systems that are more diverse. Until now, ExM has been demonstrated only for a limited number of systems, such as human, mouse, monkey, zebrafish (Freifeld et al, 2017), Escherichia coli (Zhang et al, 2016) and Drosophila (Cahoon et al, 2017). ExM needs to be applied to systems that are more diverse and further optimized based on the digestion or gelation protocol required for a specific system.…”