Drosophila ringer solution

Dissect testes from 1-day-old adult males in Drosophila Ringers Solution. 

Prepare the specimen in PBS or Drosophila Ringer solution (pH 7.2). The prepared tissue can be kept in saline until enough tissue is collected. Then, all tissue can be transferred to a microcentrifuge tube see Note 7) containing the fixative under the fume hood. Preferably, the tissue can also be directly stored in fixative (under the fume hood ) on ice until the needed number of samples is reached (rrrNote 8). 

Several buffers are suitable for dissection. Three commonly used buffers (lx PBS, Drosophila Ringer s solution, and 0.1 M sodium phosphate buffer) are given below ... 

Dissect eye discs or pupal eyes in Drosophila Ringer s solution as described in Protocol 12.1. Alternatively, perform the dissection directly in the diluted solution of acridine orange (see step 2, below). 

Incubate tissue for 5 minutes in a 1.6 x 10 M solution of acridine orange (Aldrich) in Drosophila Ringer s solution. Immediately before use, dilute 1.6 xl of a 1 mM stock in 1 ml of Ringer s. 

Mount in Drosophila Ringer s solution and cover with a coverslip. 

Drosophila Ringer s Solution (from Ashburner 1989b) 182 niM KCl 46 mM NaCl 3 mM CaClj 10 mM Tris-HCl... 

Dissect ovaries out of adult females, in Drosophila Ringer s Solution or PBS. Separate ovarioles slightly (with a tungsten needle or forceps), but maintain connections to the ovaries. 

---