Visualizing Larval Nematodes via Confocal Microscopy

Worms were on mounted on 2% agarose pads containing 120 mm Optiprep (Sigma Millipore) to reduce refractive index mismatch (Boothe et al. 2017 (link)) on premium microscope slides Superfrost (Thermo Fisher Scientific), and a 22 × 22–1 microscope cover glass (Fisher Scientific) was placed on top of the agarose pad. Worms were anesthetized using a drop of 150 mm sodium azide (Sigma Millipore) with 120 mm Optiprep. Z-stack confocal images of 24 hour old larvae were taken on a Zeiss LSM 880 microscope using a 63X objective lens.

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Kumar S., Olson A.C, & Koelle M.R. (2021). The neural G protein Gαo tagged with GFP at an internal loop is functional in Caenorhabditis elegans. G3: Genes|Genomes|Genetics, 11(8), jkab167.

Publication 2021

Optiprep Superfrost 22 × 22–1 microscope cover glass sodium azide LSM 880 microscope

Agarose Larvae Lens Microscope Sodium azide Worms

Corresponding Organization : Yale University

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Variable analysis

independent variables

Optiprep concentration (120 mm)

dependent variables

Confocal images of 24 hour old larvae

control variables

Agarose pad concentration (2%)
Sodium azide concentration (150 mm)
Optiprep concentration (120 mm) in sodium azide solution
Microscope slide type (premium Superfrost)
Microscope cover glass size (22 × 22–1)
Microscope objective lens (63X)
Microscope type (Zeiss LSM 880)

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