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Equipment/Supplies (2). We use this type of modelling clay for fixing bees. Available from most hobby stores. Equipment/Supplies (2). We use this type of modelling clay for fixing bees. Available from...
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Equipment/Supplies (4). We use Drummond wiretrol capillary tubes. Each 100 tubes come with one plunger (with the white handle). The tubes are marked for 1-5 microliters. However, because we bake all...
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Equipment/Supplies (3). We use this type of acetonitrile from EM Science. Any brand would work as long as they are HPLC grade or higher. Equipment/Supplies (3). We use this type of acetonitrile from...
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Equipment/Supplies (5) We use 12 x 125 mm cultures like this. The most important thing is that the lids MUST be Teflon lined. Teflon is resistant to chemical solvents and also JH is less likely to...
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Equipment/Supplies (6): You need a compound scope for pricking the hole and drawing blood. You do not need a high quality one (as shown here), simply 5-7 x magnification to see clearly. I have used...
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Put 500 ul of acetonitrile into each tube and cap the tube tightly. Put 500 ul of acetonitrile into each tube and cap the tube tightly.
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Clearly label each tube before the blood is put in. Otherwise you will easily forget to label it after blood is put in and your sample is wasted. Clearly label each tube before the blood is put in....
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We use an insect vacuum to collect bees from a colony. This photo shows the entrance is blocked with a piece of hardware cloth and the vacuum is used to collect returning foragers. We use an insect...
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The canister containing bees can be removed. The canister containing bees can be removed.
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Bleed the air out of the ziplock bag carefully and force the bees to form a monolayer so that they can not form a cluster inside the bag. Bees will sleep after 4-10 minutes in a refridgerator or on...
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Bees are transferred from the vacuum canister into a ziplock bag. Bees will not be anesthetized by cold if left in the canister because bees will cluster and produce enough heat for a few hours. Bees...
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Fix individual bees like this with modelling clay. Bees will wake up shortly but are unable to escape. Fix individual bees like this with modelling clay. Bees will wake up shortly but are unable to...
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You can fix a few bees at a time like this. Sometimes I fix 10 bees first, then bleed one by one. But be careful to avoid being stung by the bees -- whether bled or not, they will sting when you touch...
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This photo shows that I was fixing 11 bees and trying to get 10 samples. This was during a visit at UC Davis, when I had to bleed about 100 bees per day. This photo was taken after the bees were bled....
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After you fix the bees, your fingers will be stained with some clay. Use 70% alcohol (ethanol) to clean your fingers. Clean the plunger periodically and between the experimental groups. After you fix...
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Apply the capillary tube between the intersegmental membrane where the hole was made, hemolymph (colorless or slightly yellow-tinted) will flow out. If not, apply some pressure using the index finger...
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This step is done under a dissecting scope. Using the bent insect pin to prick a hole in the intersegmental membrane. The insect pin should be as parallel as possible with the abdomen axis, so that it...
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Use a good ruler (the finer the lines, the better), measure the millimeter length (to the nearest 0.5 mm) of blood inside the tube. This samples has 22 mm. WRITE it down. You will forget in 5 secs if...
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Stick the capillary tube into the test tube about this depth. Stick the capillary tube into the test tube about this depth.
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Insert the wiretrol plunger into the tube carefully, move the plunger down all the way to push the bee blood out, then up and down a few times. Insert the wiretrol plunger into the tube carefully,...
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