Light-microscopy methods in C. elegans research

A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults

“…This work introduces the advantages of LSFM live imaging to long term postembryonic C. elegans development, including faster acquisition speed and reduced phototoxicity and photobleaching. Prior to the development of this protocol, light-sheeting imaging of C. elegans had been limited to embryos, very short timelapse imaging of larvae and adults, and fixed samples (Chardès et al 2014; Duncan et al 2019; Chen et al 2014; Breimann et al 2019; Liu et al 2018). We anticipate that adult or larval encasement in BIO-133 within an FEP tube will enable continuous LSFM imaging for at least 2 hours, a time span that is comparable to that typical of confocal timelapses (Kelley et al 2017) and which approaches the physiological limit imposed by starvation (Schindler and Sherwood 2014).…”

phiC31 integrase for recombination mediated single copy insertion and genome manipulation inC. elegans

“…Quantitative analysis of fluorescence protein amounts has long been employed to monitor gene expression and regulation in C. elegans (B reimann 2019). While initial approaches were largely limited to detecting the presence or absence of expression by location or time (M erritt et al 2008), modern analysis techniques have become more sophisticated.…”

Probing the Functional Role of Physical Motion in Development

“…The most common technology remains the light microscope (Figure 2A), which has been used for decades to observe, describe, and quantify development. Modern microscopes are highly specialized instruments that are tailored to the needs of the biological model system (Huisken et al, 2004;Breimann et al, 2019;Jambor et al, 2015), and when complemented by suitable labeling approaches, even protein age can be measured (Durrieu et al, 2018). To quantify modes of transport in vivo, passive observations are sometimes insufficient, and fluorescent recovery after photobleaching (FRAP, Figure 2B) and photoactivation ( Figure 2C) have frequently been used to measure diffusion constants and flow rates (Firmino et al, 2013;Griffin et al, 2011;Lippincott-Schwartz et al, 2001;Goehring et al, 2011;Kicheva et al, 2007).…”