Publications and Research

Document Type

Article

Publication Date

8-9-2018

Abstract

Numerous microfluidic systems have been developed and used for live imaging of Caenorhabditis nematodes (Allen et al., 2008; Zhang et al., 2008; Krajniak and Lu, 2010; Krajniak et al., 2013; Cornaglia et al., 2015). These systems can be costly, complex to set up, or require high-maintenance between uses. In addition, microfluidic rigs can be thick, preventing live imaging of worms from strains expressing low fluorescence fusion proteins. In the absence of elaborate microfluidic rigs, most live imaging protocols utilize flat agarose pads along with anesthetics and/or microbeads to immobilize the nematodes (Kim et al., 2013). Since this method does not allow the user to maintain the nematode straight and does not prevent small movements that disturb live imaging, a higher number of worms need to be mounted to ensure that a some settle in an optimal position. This is especially problematic when trying to image nematodes genotypes that are scarce, since there is a very small number of nematodes with the desired genotype in a plate making it challenging to find enough animals to image. Here is a protocol, modified from Zhang, M. et al., 2008, to make grooved agarose pads utilizing a 12-inch vinyl Long Play (LP) record as a mold for agar pads in which nematodes can be positioned and immobilized for live imaging. This method is simple, effective, and allows long-term time-lapse imaging of young adult and adult hermaphrodites, and males expressing low fluorescence fusion proteins.

Comments

This work was originally published in microPublication Biology, available at https://doi.org/10.17912/QG0J-VT85

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Included in

Biology Commons

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.