Chapter 7 Mass Isolation of Fly Tissues

Abstract

Publisher Summary This chapter discusses the techniques for isolation of biochemical quantities of Drosophila cells. One of the principal limitations in the use of Drosophila has been the difficulty in obtaining sufficient quantities of cellular material from an organism that weighs only 1.5 mg and an egg weighing approximately 0.01 mg. Although many protocols are available for mass rearing of flies, the methods described in this chapter are developed because of the need for having large quantities of embryos. Using the protocol, 300 gm of viable eggs are produced daily and a kilogram of adult flies weekly. To maintain population cages of flies producing many grams of eggs and other stages of Drosophila development, a protocol is developed that eliminates the requirement for typical cooked fly food. This approach utilizes a liquid yeast broth, poured onto an absorbant cotton matrix, on which 1 ml of eggs are pipetted. By using dead yeast, populations of flies could be maintained with sufficiently low levels of contaminating yeast so that sterile cell cultures could be readily established later.