Frequently Asked Questions

Check a local gas or beer supply store. Tanks are usually available for rent with a refillable policy. You also need a regulator; these are specific to the gas. Use a 2 stage CO2 gas regulator as this will allow fine control of the flow rate to a slow bubble.

A magnification 7X to 20X (most go higher). Note: less magnification is needed for good depth of field. An extra 0.5 lens will reduce magnification. A working distance 4.5 " or 110 mm. is necessary. The type of microscope stand and working distance is critical to accommodate the Instrument. A flat base with a width of 7" or 18 cm, or a boom stand is recommended.

Use a gooseneck LED light. This provides flexibility in adjusting and directing the light around the instrument. The LED is a cool light, avoiding heating and drying of delicate tissues. Note, a microscope bottom light is unnecessary and most of these are detachable.

A flow rate of 35ml / minute is recommended. With a slow steady bubble, the queen should be motionless in about 15 sec. Note, queens do have different tolerances, so this can vary between queens. Adjust the flow before inseminations. A wash bottle or flow meter can be used to adjust and visualize the flow rate. Neither is essential if you use a two-stage CO2 regulator. The two-stage regulator allows fine flow adjustments to a slow bubble.

Two CO2 treatments are recommended to initiate egg laying of IIQs within 3 -6 days after insemination. A brief, initial treatment a day before insemination is recommended vs after. As, this does not affect sperm migration and generally initiates oviposition earlier than giving a treatment after insemination. The second CO2 treatment is given during the insemination procedure.

CO2 treatments should be brief, just enough until the queens become motionless. For the initial dose , a day before the insemination procedure – Several or a frame of caged virgins can be placed in a plastic bag and gassed. Given a slow bubble, this can take a few minutes, 2 to 4 minutes is optimal.

A single large dose of is 8 to 10µL or two small doses  of 3 to 5µL are recommended. The standard semen dosage is 8 to 10µL. One dose of 10µL is recommended, as IIQs tend to be productive for an additional 6 to 8 months, vs a dose of 8µL Or, two small doses can be given a day or two apart. Smaller semen doses migrate faster and may help avoid plugged oviducts due to poor sperm quality or poor queen care conditions.

Many factors can contribute to delayed oviposition; infection, injury, plugged oviducts, poor post insemination care of queens, seasonal effects, inadequate nutrition, temperature extremes, etc. Optimize insemination procedures and beekeeping practices.  If queens look healthy and active, an additional brief CO2 treatment can be given to stimulate egg laying. If egg laying is not initiated in a few days, the cause could be due serious factors as plugged oviducts, a slight injury or infection and these queens usually are killed or remain poor layers. Generally, injury or infection will cause IIQs to become lethargic and die. Spermathecas can be dissected to diagnose if the oviducts are plugged (congealed appearance), or infected (discolored). See the I.I. video for details, photos.

Drones are fragile, highly susceptible to adverse and stressful conditions. Many factors play a role in their quality and numbers.  Poor nutrition (especially

during development), chemical residues (miticides and agricultural), and exposure to temperature extremes will impact sperm numbers and quality. As you need a plentiful supply of mature, well-fed , well cared for drones, anticipate many of these will be unusable due to lack of semen and contamination during the explosive eversion process. Sometimes simply improving your techniques, proper and efficient eversion the endophallus, will increase success. During semen collection, keep drones warm and fed, as they become lethargic after about 30 min. without proper care and become more difficult to evert.