3.1.3. Reagents

  • Alcohol has a variety of uses. The stock should be 95 % Industrial Denatured Alcohol (IDA). The best dissecting fluid is 30 or 50 %, made by diluting the stock with water (30 parts spirit to 65 parts water by volume, or 50 to 45, respectively). Water cannot be used by itself because it will not wet the hairy bodies of bees.
  • Formalin (38-40 % solution of formaldehyde) will be needed as preserving fluid. Caution: toxic compound.
  • Bees can be killed with many volatile substances such as ethyl acetate or chloroform, but freezing is also a good method of killing bees (see Human et al., 2013).
  • Glacial acetic acid will be required if Carl's solution is preferred for preservation; it has many advantages (see section 3.1.4.).
  • Glycerol, diluted with water, is an excellent preserving fluid for finished dissections.
  • Insect saline solution is a good preservative for dissections: 7.5 g NaCl, 2.38 g Na2HPO4, 2.72 g KH2PO4 in 1 l of distilled water. Insect saline is preferable to alcohol as it preserves the tissues for the time of dissection, but it does not harden the tissues as alcohol does.
  • Either saline solution or Grace’s insect tissue culture medium should be used when observing organs in vivo. These solutions are isotonic relative to the organisms body fluid. Sterile bee saline is appropriate for honey bees and can be easily made. Sterile bee saline is composed of 130 mM NaCl, 6 mM KCl, 4 mM MgCl2, 5 mM CaCl2, 160 mM sucrose, 25 mM glucose, 10 mM 4-(2-hydroxyethyl)-1-piperazineethane- sulfonic acid in distilled water (pH 6.7, 500 mOsmol) (Richard et al., 2008). Alternatively, Grace’s Insect Tissue culture medium can be purchased from various biological supply companies or made according to Grace (1962).