Drosophila facility information

General information

• Fly Lab D22 19°C (0114 22) 22811
• Media Room D34a (0114 22) 32802
• Culture Rooms: D22b 18°C, 70%RH; D22c 25°C, 70%RH

Fly facility technicians

• Kath Whitley (Fly Facility Manager) K.Whitley@sheffield.ac.uk
• General fly technician email flytechnicians-group@sheffield.ac.uk

Other useful contacts

• Martin Zeidler m.zeidler@sheffield.ac.uk (fly academic lead)
• Melanie Hannah m.hannah@sheffield.ac.uk (School Technical Manager)

The risk assessments and CoSHH forms for the fly lab and media room are in labelled blue files in the relevant rooms.

General fly lab rules

If the CO₂ sensor is sounding then this indicates dangerous levels of CO_2. Leave the lab immediately and prop open the door. 

Please ensure you turn off your CO₂ if you are not using it and after you have finished using your fly station.

No food or drink is to be consumed in the laboratory.

When listening to music via headphones please keep the volume down so that others cannot hear it and to ensure you can still hear any alarms that may sound.

Please swab down your bench with 70% IMS after you have finished your fly work each day.

When you have finished using your microscope make sure the light source is switched off.

If you find any mites please tell Kath so the problem can be dealt with immediately.

Only keep fly cultures for the maximum time indicated:

• 18°C controlled temperature room – vials/ICRF bottles 6 weeks; no straight sided bottles
• Main lab – vials/ICRF bottles 6 weeks; Straight sided bottles 4 weeks
• 25°C controlled temperature room – vials/ICRF bottles/Straight sided bottles 3 weeks

If the fly food in the main lab is running low it is your responsibility to get more from the cold room D33.

Please ensure you label all trays with your name and all bottles with the date.  Trays of vials tipped together can be dated on the front of the tray or the first vial in the row.  All vials bundled together by elastic bands should be dated individually.

Ensure you fully close controlled temperature room doors (the light should not flash).

Keep a record of your fly food use (either individually or on whole lab record).

Media and equipment

Fly media is stored in the cold room D33.  If you need to use small quantities of food, some will be available in D22 (on the long bench next to the door).  If this is running out or you need larger quantities please fetch more from the cold room.  Please use the earliest dated (ie oldest) food first.  If you require large amounts of media for an experiment please co-ordinate with Kath at least two weeks before you wish to use it. 

Apple Juice plates are available in 60mm sizes.  These are stored in the cold room and can be put into the fridge in D22 as needed.  If you would like 90mm or 100mm apple juice plates please ask one of the fly lab technicians the week before you need to use them.

Please put all used trays next to the food on the long bench for checking and washing by fly technicians.  Put all small ties in the labelled beaker so they can be reused. 

Please wash up any egg laying pots and re-usable yeast containers.

Spare equipment such as paintbrushes and light bulbs for microscopes are available from one of the fly lab technicians.  Please ask if you can’t find an item you require.

Fly stock collections

Remember much of the stock collection is irreplaceable. It is very important to avoid contaminating or otherwise harming stocks.

Please always get permission from the relevant Principal Investigator before taking stocks – this may also ensure you get the stock you really need!

Do not deplete the flies from a stock, to the extent that it threatens the health of the stock. In general, it is OK to take flies from the back vial, provided the other vials are healthy (you should check this!). Only take flies from other vials with great care.

Do not remove stocks from the collection, except briefly when collecting flies and always return promptly. Be very sure to return stock to the correct position in the tray.

Any flies that are obtained from sources outside the laboratory must be quarantined before bringing them into the fly lab to make sure they are not carrying anything that can spread to other stocks. Allow them to go through 2 generations and examine the culture carefully under a microscope to check for mites before bringing them into the fly lab or culture rooms.  Mites can out compete weak fly stocks or even kill them so please be vigilant and tell Kath if you see old cultures that need to be dealt with or if you discover any mites.  Then we can stop them spreading.

Adult mites are smaller than fly embryos and often walk on the sides of vials and bottles around the pupae and empty pupal cases.  Mite eggs and adults can be observed in Drosophila culture using the youtube link below.

 https://www.youtube.com/watch?v=PfSAAsrVANc#action=share

If you know your cultures contain mites and you don’t need them, freeze the vials overnight to kill the mites.  Place the frozen cultures into a yellow waste bin.  Also freeze any fly cultures from outside sources before disposing of them.

If your culture contains mites and you wish to keep the flies, remove cultures from the fly lab. Passage them three times on consecutive days onto fresh medium.  Then allow the flies to go through two generations.  Putting the tubes on a surface of glycerol makes the transfer of mites from one tube to another more difficult.

Waste

Please refer to the waste assessment and poster by the door in D22 for information regarding the correct segregation and disposal of waste.

Use the yellow bins to dispose of fly cultures.  These will contain an orange bag on the outside and two black bags on the inside.  When the bin you are using is full please take out the bags and twist the top around.  Secure very tightly with a long electrical cable tie ensuring no flies can escape.  Fill out an orange label (GM/Infectious Waste) with the following information: Department-BMS Laboratory–D22 and Date Sealed.  Place the label on the bag and put it in the designated place for disposal.  

Put an orange bag and then two black bags into the yellow bin.  

The sharps bin is located on the long bench.

Use the bin with black lid for non-hazardous waste and the bin with green lid for recycling.

Rinse used Fisher Winchesters and dispose of the lid.  Deface the label with a cross and place the Winchester outside the lab for collection.

Carbon dioxide

If the carbon dioxide alarm is sounding:

Do not enter the fly lab.  Wedge door open to ventilate lab.

Inform either:

• In Emergency Internal 4444, mobile 0114 22224444
• Out of hours The University Control Room 0114 2224085

If you are in the fly lab when the carbon dioxide alarm is sounding:

Ensure your CO₂ tap is turned off.

Exit the lab and ensure there is nobody in the lab.

Wedge the door open to disperse the CO₂.

Then call numbers as above.

Note: If the alarm is flashing but not sounding the CO₂ will have exceeded the 8 hour safe long term exposure limit (0.5%).  Do not work in the lab for extended periods under these conditions.  Do not wedge the door open as this breaches GM regulations.

Please remember to turn off your CO₂ tap when you have finished using it.

If you suspect there is a CO₂ leak inform Kath Whitley.  Out of hours contact the University Control Room.

CO₂ supplies can be turned off to each bench in the fly lab by closing the black valves underneath the benches.

Culture room alarms (D22b and D22c)

The alarm will go off if the temperature or humidity goes above or below limits or there is a power failure.  If there is a temperature fault please open the culture room door until the fault is rectified.  Ask Kath Whitley to contact the EFM helpdesk or out of hours contact University Control Room on (0114 22) 24085.
 
If the autodialler rings the fly lab phone nearest to the door to report a fault.  Press 8 on the phone to cancel the message.  If you do not get to the phone in time please enter the default code 1234 on the alarm panel to stop it phoning around everyone.

D22 Air Conditioning

The fly lab temperature should read 19°C on the thermometers above Kath Whitley and David Strutt’s work stations.  If the temperature on the thermometers reads below 17°C or above 20°C inform Kath Whitley immediately.