Visualizing and quantifying molecular and cellular processes in Caenorhabditis elegans using light microscopy

Abstract

Light microscopes are the cell and developmental biologists’ “best friend,” providing a means to see structures and follow dynamics from the protein to the organism level. A huge advantage of Caenorhabditis elegans as a model organism is its transparency, which coupled with its small size means that nearly every biological process can be observed and measured with the appropriate probe and light microscope. Continuous improvement in microscope technologies along with novel genome editing techniques to create transgenic probes have facilitated the development and implementation of a dizzying array of methods for imaging worm embryos, larvae, and adults. In this review, we provide an overview of the molecular and cellular processes that can be visualized in living worms using light microscopy. A partial inventory of fluorescent probes and techniques successfully used in worms to image the dynamics of cells, organelles, DNA, and protein localization and activity is followed by a practical guide to choosing between various imaging modalities, including widefield, confocal, lightsheet, and structured illumination microscopy. Finally, we discuss the available tools and approaches, including machine learning, for quantitative image analysis tasks, such as colocalization, segmentation, object tracking, and lineage tracing. Hopefully, this review will inspire worm researchers who have not yet imaged their worms to begin, and push those who are imaging to go faster, finer, and longer.

The role of Caenorhabditis elegans in the discovery of natural products for healthy aging

“…215,216 The worms' transparent body allows for researchers to visualize and quantify molecular and cellular processes like the expression, localization and activity of proteins. 217 A frequently employed approach is to use transgene worms carrying uorescent protein tagged transcription factors to test for nuclear translocation in response to treatment. 218 In this case daf-16::gfp (e.g., strain TJ356) and skn-1::gfp (e.g.…”