First we gathered red roses (Rosa berberifolia) from Betty's garden, tulips (Tulipa gesneriana) from Mrs. Lindahl's collection, and winter pansies (Viola tricolor) from Mrs. Lindahl's collection. To prepare the roses, we put them in the dehyrator for 48 hours to dry them out because the SEM cannot have any moisture inside of it. The tulips and pansies were already dried out.
Rose
Winter Pansy
Step Two:
Then we loaded the stub that we'll use in the SEM microscope. We took the stub and peeled off the white tape. We pushed the stub on to a piece of carbon tape that was on a sheet, then peeled the stub (attached to the carbon tape) off of the carbon shape.
We gathered all of our samples--the roses, tulips, and winter pansies. For each sample we had a different paintbrush. One at a time, we used the appropriate paintbrush to brush a sample of pollen onto the stub.There were three different sections on the stub, one for each of the pollen samples.
When we had finished loading the pollen samples on the stub, we blew off the stub with a multi-purpose duster, so that loose pollen wouldn't come off in the SEM machine. Then the stub samples were prepared, so we put the stub into a small white box labeled with our names for storage.
Step Three:
We prepared pollen samples to look at under the microscope. We took brushes and brushed pollen from each flower onto a glass slide. Then we added a drop of water, and put a cover slip over it. Under a Swift Ultra microscope, we looked at each sample under 400x. Unfortunately, we couldn't find any pollen from the winter pansy, but we got photos of the rose's and tulip's pollen.
Tulip pollen, 400x
Rose pollen, 400x
Step Four:
Next we looked at our samples under a Leica EZ4 HD microscope, using Leica Imaging 1.0. We looked at our prepared stub, and our original flower collections that were in bags. We looked at the winter pansy, tulips and roses one at a time under the microscope. Unfortunately, our Leica photos were never loaded off of the computer before the computer was taken away, so we don't have those photos. However, we have comparable photos taken on a Frey Scientific microscope at 35x.
Rose, 35x
Red tulip, 35x
Winter pansy, 35x
Step Five:
Now we used the SEM (Scanning Electron Microscope) to take pictures of our samples. First, we picked up the stub with two fingers from the storage box. We took the cup from the SEM, and pushed the stub down into the cup. We lowered the sample to be even with the cup (we eyed it), by turning the silver top part. Then we used the side notch on the cup, and clicked over four times to the right, which makes sure that the sample is low enough to go into the SEM. We placed the cup into the slot of the SEM, and pushed all the way in, until the green light came on. Then we pushed down the door of the SEM.
On the screen, we pressed the scanning button. We mapped the entire sample by pressing the button that looked like a maze. Then we went into settings and pressed label to label our sample, then clicked okay, then went back to the image. We started scanning our sample, and when it was ready we moved around it and took pictures. Here are the pictures we took below.
Tulip Pollen Pictures
Collection of several pollen grains, 360x.
Pollen grain, 2720x.
Texture of pollen grain, 5050x
Pollen grain, including the length measurement, 2000x.
Rose Pollen Pictures
Collection of pollen grains, 2600x.
Grain of pollen, 4200x.
Grain of pollen that shows its three apertures, 6500x.
Same grain of pollen as above, 12000x.
Grain of pollen that shows the measurement of the length of the grain (25.6 um)
2000x.
Same pollen grain as above, displays the length and width measurement of the grain (45 um long) 2000x
Winter Pansy Pictures
One grain of pollen, showing an aperture, 725x
Photo from http://pollenlab.blogspot.com
Pollen grain with decollate apertures, shows length measurement of grain (75 um), 2000x
Photo from http://pollenlab.blogspot.com
Photo from http://pollenlab.blogspot.com
Step Six:
The first thing that we did was we looked at all the background that we currently had and we decided that what the two closes relating flowers would have to be. With that information we made our pollen morphology.
Step Seven:
What we did was find out what the names of our flowers were and we went on a web sit that gave us information about our flowers. Then we found out which flowers were really related because of what flowers had the most proteins they had in common. Finally we made a tree of our flowers by protein sequence.
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