Procedure

1. Preparation for the SEM:

To prepare a pollen sample of the Lilium asiatica (Asian Lily), we first cut the anthers of three different lilies within the bouquet. Once dehydrated, we prepared the samples for the SEM. First, we placed a piece of carbon tape on the stub. Then, we used a different brush for each pollen sample to place a small amount of pollen onto the stub. After that, we set the sample in the tray. Before placing the sample into the SEM, we used liquid air to blow off any loose pollen--moving from low to high pressure. Once this was completed, we placed the sample in the cup. The next step was crucial: we clicked the top notch of the SEM four times to the right. Then we placed the cup into the slot and once the green light turned on, we pulled down the door. We devoted the remainder of our time to mapping specific areas of the sample and taking measurements. 

2. LEICA Procedure: 

Stage 1 Asian Lily (35X)

Stage 2 Asian Lily (35X)

Stage 3 Asian Lily (35X)

We used the Leica EZ4D Microscope to obtain images of our pollen at a magnification of 35X. With its 7 LEDS, the Leica is a microscope that provided us with high quality illumination and contrast in the images of our pollen samples.








After putting small samples of pollen at each stage in separate dishes, we placed it under the microscope. Three people were needed to operate the Leica and obtain the images of our sample. Two were needed to adjust and focus the lens onto the specimen and one was needed to capture the images at the right moment. We experimented by adjusting the light at various angles to optimize our view of the sample.







Unfortunately, we were unable to recognize any noticeable differences between the pollen grains at their various stages. We expected to see a greater distinguishing characteristics at the SEM.

Julia preparing the sample for the Leica. 

3. Compound Microscope:

At Station 8, we created wet mounts of the pollen samples we had left over of our Asian Lilies. We used the growth media solution in place of water. The image below is the pollen of our Asian Lily at Stage 2.

Asian Lily Stage 2 (400X)

 From the image, we can see one aperture. There are also distinguishable features on the surface of the pollen grains.

Christina observing the pollen sample!

4. SEM Work:

While working with the SEM Phenom Pro by FEI, we were looking between fiduciary marks to focus on different stages of our pollen samples. After many adjustments, we worked to focus on individual pollen grains and captured their image at a magnification of 2000X. (See SEM photos on sidebar.) 

Kent focusing on SEM images!