Worm injections with SPD-2

As PCR of SPD-2 showed the cleanest band of all four genes, we proceeded with worm injections with SPD-2. After making the injection mix and checking that the worms were ready for injection, my supervisor injected the SPD-2 mix into the worms.

This step was carried out by my supervisor as these injections require a lot of practice and is highly intricate work. However, it was interesting to see they set up for the injections and how precise everything needed to be to get the injection mix into these worms.

Injection apparatus fixed upon a microscope

I was shown how the needles for the injection are made, using the machine (shown below) and glass capillary tubes. The tubes are heated in the center, and eventually the weight pulls the capillary tubes apart, leaving a very fine glass needle. This needle is then what will be used dto inject the worms with. These needles are required to be small given C. elegans grows up to 1mm in length - so a fine needle is required to ensure an accurate injection.

Machine used to make fine needles for injection.

I was also shown how the needle was loaded with the injection mix, and how the worms were mounted onto a slide, using agarose and mineral oil to prevent the worm from moving around as well as drying out.

The worms were then injected with the SPD-2 and progeny of the injected worms were picked onto seperate plates for screening.