Pollinator collection and pollen load analysis (this is Amanda!)

We have made a few changes to the protocol for tomorrow’s FINAL outing for pollinator observation and collection:
1) DO NOT capture syrphid flies!
2) If you observe a small common syrphid fly (Family: Syrphidae, Sphaerophoria sp.) on your Echinacea head, make a note that one SCSF was observed. Here are some photos of syrphid flies:

http://echinacea.umn.edu/insects/images/poll2005vin1948side.jpg
http://echinacea.umn.edu/insects/images/poll2005vin1927top.jpg
http://echinacea.umn.edu/insects/images/poll2005vin1927front.jpg

Here’s a video that might help:
http://www.youtube.com/watch?v=UkjPfAMGmiw&NR=1

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As for pollinator collection and analysis…

We’ve gone out collecting three times now (tomorrow is the fourth and final day), and we have about 140 Echinacea insect visitors in vials in the freezer. Here is my protocol for making slides and labeling specimens:
1) Remove vial from the freezer and allow insect to thaw
2) Cut a very small (2mm cubed) piece of agar and situate it in the middle of a slide.
3) Remove the insect from the tube and dab and sweep every surface of the body across the agar cube. This is intended to simulate the amount of pollen that might be transferred to Echinacea styles during a visit.
4) Set the insect aside on Styrofoam.
5) Sweep the inside of the vial with the agar cube for any loose pollen.
6) Place a cover slip atop the agar cube and put the slide on the edge of a hotplate on LOW heat. Watch the slide carefully and remove it when the agar is soft but not melted (once melting starts, it will quickly boil and create bubbles in the slide).
7) Pin the insect (for identification if it is an unknown species, or quick-n-casual for a known species) and pin it with a new one letter, three number code.
8) Label the slide with the vial code and the specimen code.

Photos:
I photograph the slides on the same day as I make them so that the agar doesn’t dry out before the photo. To sample the slides, I have generated random pairs of numbers from 1 to 22 (inclusive). The cover slips are 22 x 22 mm and I use the ruler on the microscope to scroll to the randomly selected “plots” from one corner of the cover slip. From there, I take the photo at 40x. I take ten photos for each slide, and sometimes take more than one photo per plot if I capturing all of the pollen grains requires multiple focuses.

I will post some photos of this process soon, so hang in there!
As always, feel free to leave questions or comments.

Thanks!!
Amanda