Protocols

Hanging drop method for C. elegans embryos

Introduction

In this protocol, we show how to prepare C. elegans embryos on a standard coverslip to perform experiments with CherryTemp using the hanging drop method. It shows step by step how to dissect and mount the sample for the experiment.

Imaging Drosophila third instar larval brain cells expressing Jupiter-mCherry as a microtubule marker

Step-by-step guidelines:

Step 1: Pick young adults into pre-cooled M9 medium (16°C).

methods-c-elegans-embryo-temperature-control-agar-pad1-e1530786207296

Step 2: Dissect them by cutting at both ends of the uterus using a scalpel and #15 blade to release the embryos.

methods-c-elegans-embryo-temperature-control-agar-pad2bis-e1530786233192

Step 3:

Choose the embryos of the desired stage and transfer them to a cover slip using a mouth pipette.

methods-c-elegans-embryo-temperature-control-agar-pad3-7

Step 4:

Use a syringe to make a ring of Vaseline on a coverslip. Make sure to center the ring so that the embryos will be correctly placed under the thermalisation pattern of our chip.

 
Hanging-drop-protocol-for-C.-elegans_Fig_All-01-e1530794234483

Step 5:

Use a mouth pipette to transfer the embryos, cluster embryos with an eyelash tool.

Hanging-drop-protocol-for-C.-elegans_Fig_All-02-e1530794279894

Step 6:

Carefully cover with our temperature control chip. Gently clamp the system using our insert.

Hanging-drop-protocol-for-C.-elegans_Fig_All-03-e1530794311474

Step 7:

Place the mounted slide on either an upright or an inverted configuration by simply rotating 180°.

Hanging-drop-protocol-for-C.-elegans_Fig_All-04-e1530794342825

References

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