Drosophila Mannelijke accessoire klier dissectie: een methode om secundaire cellen te isoleren

- Under a dissection microscope and in serum supplemented media, remove the reproductive tract from each fly with forceps. First, remove parts of the digestive tract that may have accompanied the reproductive tract. To isolate the accessory glands and ejaculaory duct, remove the testes, the ejaculatory bulb, and the terminalia. Wash the tissue for one to two minutes in PBS to remove the serum. Then, transfer it to dissociation solution.

To expose the cells inside the glands to the dissociation solution, hold the center of each gland with forceps and slice through the gland with a pair of sharp forceps. Remove the proximal portion of the gland and the ejaculatory duct to leave behind the distal end of the glands containing the rare secondary cell population responsible for secreting proteins involved in mating. Finally, transfer the accessory gland tips to a micro centrifuge tube in preparation for digestion. In the following protocol, we will dissect the accessory glands to isolate secondary cells for transcriptome analysis.

- Before beginning the procedure, cut and flame round, wide 200 microliter tips for handling the glands, and pass the tip opening of multiple 1000 microliter t