Nematode Slide Preparation: A Method to Mount Animals on an Agar Pad

– First, place a microscope slide between two other slides with tape. These provide support and spacers for the agarose pad preparation. Next, place a drop of melted agarose onto the slide. Immediately press down on it with another slide.

After the agarose solidifies, carefully separate the slides so that the agarose remains on one slide. Always use freshly made pads as dried out pads can dehydrate the animals. Ensure there are no cracks or bubbles, which can disrupt pad integrity.

Next, add a drop of anesthetic para the pad center and transfer 10 para 15 animals para it, bringing as little bacteria with them as possible. Excess bacteria could interfere with imaging. If the anesthetic solution dries out, add a second drop onto the animals.

Gently place a coverslip over the pad. Avoid forming bubbles or placing too much pressure on it para prevent damaging the animals. Finally, label the slide, which is ready for imaging.

– Start by preparing slides for imaging. Place a drop of melted agarose onto a microscope slide and immediately press down the droplet with another microscope slide gently flattening the agarose Need the agarose para dry for one minute.

Then, carefully separate the two slides and leave the solidified pad on one of the slides. Place the slide on the microscope stage and add a 5- para 10-microliter drop of anesthetic para the center of the agras pad.

Use a heat sterilized pick para quickly transfer 10 para 15 animals from the stock plate para the anesthetic droplet before the solution dries out. Then, apply a coverslip by positioning it just above the agarose pad and gently dropping it down.