The basic technique of I.I. has not changed significantly since its development in the 1950s. Proficiency requires practice, precision, and sanitary conditions. Specialized beekeeping skills and proper care of queens and drones are essential to quality control. Several options of instrumentation are currently available, which offer choice but can vary in quality and lack standardization. The basic instrument consists if a stand, a set of hooks, queen holder assembly, syringe, and syringe tips (Figs. 1, 2, and 3). The microscope stand must be compatible with the instrument and provide sufficient depth of field and instrument clearance (Fig. 1). A cold light source is also recommended to prevent heating and drying. A source of carbon dioxide with flow regulator and flexible tubing to the instrument are also required.
Equipment requirements include:
- Complete insemination instrument, including an instrument stand, manipulators, syringe, and accessories (available through specialty honey bee supply companies)
- Binocular stereozoom microscope, 10x to 20x, and cool light source
- Carbon dioxide source with flow regulator and tubing
- Saline solution (see section 2.2.)
- Sterile vials
- Pipettes and bulb or syringes
- Distilled water
- 95% ethanol
- Sodium hypochlorite
- Sterile tissues and cotton swabs
- Squeeze bottles
- Paper towels or kimwipes
- Autoclave or pressure cooker (for sterilization)
- Queen cages
- Drone holding cages and drone flight box (Fig. 4)
cages are made of queen excluder material to allow worker bees access to care
for them when held in nursery colonies. Cages can vary in size, although they
should be sized to fit in a frame space in nursery colonies and also fit easily
in a flight box. Drones are released into the flight box for easy access during
semen collection (Fig. 4).
Fig. 1. Standard equipment and arrangement for performing instrumental insemination of honey bee queens. (A) a dissecting microscope, (B) handle for ventral hook, (C) ventral hook base (left C arrow) and ventral hook (right C arrow), (D) syringe base (upper D arrow) and syringe (lower D arrow), (E) syringe plunger, (F) plastic tubing leading to CO2 source, (G) chamber in which queen is placed, (H) sting hook (left H arrow) and sting hook base (right H arrow), (I) handle for sting hook, and (J) microscope base.
Fig. 2. Schley Instruments with micro-manipulated syringe, set of dorsal and ventral hooks, queen holder assembly with CO2 attachment (for labelled parts, see Fig. 1).
Fig. 3. Harbo large capacity syringe designed for semen collection and storage.
Fig. 4. Drone holding cage and flight box. Drones are collected in holding cages made of queen excluder material (left photograph). Cages should be sized to fit in a frame space in a nursery colony and also fit into a flight box (photograph on right: holding cage is left of white/screened flight box). Drones are released into the flight box for easy access during semen collection.