Last Updated: 06/15/2026

Procedure

Day 0: Hatching and panning

Preparing hatch jars:

1. Fill mason jars with 600 ml of Nanopure water
2. Cover loosely and add autoclave tape
3. Autoclave on Liquid-40 cycle
4. Tightly seal jars immediately after removing from autoclave
1. Note that eggs hatch best in deoxygenated water, so keeping the jars sealed before a hatch is important
5. Store hatch jar in incubator (28°C) at least 24hrs before use

Hatching mosquitos:

1. Using regular scissors (that you can clean with ethanol), cut off desired amount of egg sheet
1. (1 sheet is ~1500 eggs, 1 pan can hold ~300). Usually, the desired amount is about 1 inch/3 cm of paper.
2. Submerge egg sheet in glass jar with warm (28°C) autoclaved Nanopure water, place lid on, let soak for 15 min or until enough larvae have hatched.
3. Prepare a Tupperware container: fill halfway with warm Nanopure water (ideally 28°C)
1. Our containers typically hold ~500 mL
4. Count out 300 larvae using a pipette and place into container (~10 at a time, using a program such as https://simplecounter.app to keep track)
1. Use an appropriate pipette tip and trim the end of tip to prevent damaging the larvae
5. Feed larvae appropriate amount of food (follow figure referenced in the Food section above; 60 mg ground Tetramin), cover container with lid (tape label with date and time)

Day 1-6: Monitoring and feeding larvae

1. Drop in food once a day (reference food table above)
2. Obtain a clean Tupperware container and fill halfway with Nanopure water (28°C) (reference step 3 under “Hatching Mosquitos”)
3. Wash mosquito larvae by transferring with a micropipette to a clean container (small beaker or petri dish) filled with clean Nanopure water.
4. Transfer larvae from container of choice to Tupperware (prepped in step 2) using a micropipette. As larvae are transferred, keep track of count; you should transfer around 300 larvae.
1. During steps 3 and 4, mosquitos can be washed and transferred in groups. Monitor wash water to ensure it remains clean.
5. Monitor growth: will molt through 4 larval stages (instars)

Day 7-14: Transferring pupae to emergence jars

1. Fill bottom of emergence jars with 1in of distilled water
2. Use widened transfer pipette or scoop to move pupae from pan to emergence jar
3. Label jar with date and number of pupae transferred (max: 50 pupae)
4. Soak cotton ball with sugar water, place on top of screen, cover with petri dish
5. Re-soak cotton ball 1x per day until finished with adults

Reference Photos:

Day 0

2 Days Post Hatch

3 Days Post Hatch

4 Days Post Hatch

Day 5

Day 7

Day 8

Day 9

Day 10

Day 12

Milling L1 food

Using the Mill:

1. Pick out a mill sifter attachment out of the three labeled 20, 40, and 60
1. For mosquito ground food, use the attachment labeled 40
2. Turn clear knob holding the glass plate, to the right, to remove glass plate
3. Place sift attachment in its given slot on the mill.
1. Be sure to have the sift mesh side up.
4. Slide the blue section up to hold attachment in place.
1. Use black knob for tightening and loosening
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Reference photo on the right

5. Place glass plate back into place using the clear knob
6. Place any container or jar underneath to catch product
7. Plug in mill to power source
8. Place substance you are planning on milling in the funnel on top of the device
1. If the wooden dowel is in funnel, take out
9. Flip switch on mill to turn on

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Be sure to clean out mill appropriately when done