2020
DOI: 10.1016/j.heliyon.2020.e04263
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Abstract: Background Caenorhabditis elegans is a model organism used to study gene, protein, and cell influence on function and behavior. These studies frequently require C. elegans to be immobilized for imaging or laser ablation experiments. There are a number of known techniques for immobilizing worms, but to our knowledge, there are no comprehensive studies of the various agents in common use today. New method This study determines the… Show more

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Cited by 15 publications
(14 citation statements)
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“…Conventional anesthetic assays can have detrimental effects on animal physiology, i.e., it can alter muscle tone consequently altering body length, and cannot easily be used for long-term imaging of the same animal. Anesthetic-free immobilization using a microfluidic device shows robust vesicle transport ( Mondal et al, 2011 ), fast axonal regeneration rates ( Guo et al, 2008 ), and quick behavioral recovery ( Dong et al, 2018 ; Manjarrez and Mailler, 2020 ). In agreement with our previous vesicle transport studies ( Mondal et al, 2011 ), we found robust mitochondrial transport in the device immobilized animals.…”
Section: Discussionmentioning
confidence: 99%
“…Conventional anesthetic assays can have detrimental effects on animal physiology, i.e., it can alter muscle tone consequently altering body length, and cannot easily be used for long-term imaging of the same animal. Anesthetic-free immobilization using a microfluidic device shows robust vesicle transport ( Mondal et al, 2011 ), fast axonal regeneration rates ( Guo et al, 2008 ), and quick behavioral recovery ( Dong et al, 2018 ; Manjarrez and Mailler, 2020 ). In agreement with our previous vesicle transport studies ( Mondal et al, 2011 ), we found robust mitochondrial transport in the device immobilized animals.…”
Section: Discussionmentioning
confidence: 99%
“…To image starved animals, young adults were rinsed 3X in M9 buffer and transferred to fresh unspotted NGM plates with 200 μg/ml ampicillin (to inhibit bacterial growth) for 24 hours before imaging. Worms were mounted on 5% agarose pads and immobilized with 1 mM levamisole in M9 buffer (Manjarrez and Mailler, 2020) and imaged using a Nikon Eclipse TE2000-S inverted microscope equipped with a Plan Apo 60×/1.25 numerical aperture oil objective (Nikon Instruments Inc., Melville NY, USA). Images were captured using a Q imaging Exi Blue camera (Teledyne Photometrics, Tucson AZ, USA) and Q Capture Pro 7 software (Teledyne Photometrics, Tucson AZ, USA).…”
Section: Methodsmentioning
confidence: 99%
“…To verify DAF-16 activation we used the CRISPR allele of daf-16 tagged at the C-terminus with GFP [ 90 ]. Since DAF-16 translocation from cytoplasm to nucleus is induced by heat stress and common drugs used to anesthetize the worms [ 91 ], the young adult worms were fixed in 4% paraformaldehyde. The worms were grown at 20o C on RNAi plates, at L4 stage 15 μM FUdR was added and next day, the young adults were fixed and immediately visualized.…”
Section: Methodsmentioning
confidence: 99%