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Recap:
Since December, I've been growing my Arabidopsis plants. Soon, they'll have flower buds, and I can begin counting pollen and actually getting some data. While waiting for them to flower, my main priorities have been practicing and refining my methods, and working on writing my paper.
Determining Methods
My main priority was exploring different methods for pollen extraction and imaging, finding which one would be best for my experiment, and practicing it to minimize errors with the Arabidopsis flowers. To do this, I bought regular lilies from the grocery store to practice on.
A lily flower, partially dissected to make the anthers more visible.
Anthers were placed in a microcentrifuge tube for two days to allow them to release their pollen. Then, the tube was vortex mixed to ensure all pollen was out. The debris from the anthers was removed, and the pollen was suspended in distilled water.
Lily pollen under a microscope.
Arabidopsis flower buds.
The data I have so far.
Data Collection!
Data Collection!
Some of the plants finally started flowering, so I was able to officially begin collecting data!
My Procedure:
As described above, the pollen was immersed in 2mL of distilled water to make imaging easier. However, even 2mL is quite a lot to look at under a microscope, especially at the magnification required to view pollen. Therefore, I decided to use samples of 5uL (1mL = 1000 uL) and then multiply to get an estimate of how much pollen the total 2mL would have. For each flower, I analyzed five of these samples, then took the average of each, ensuring that my data wouldn't be as skewed by any outliers.
What's Next?
Moving forward, I need to keep collecting data and start to prepare to present my project in March/April. So far, the only plants to flower have been from the testing group with the modification, so I can't start comparing the data quite yet. However, even more plants are beginning to show buds, so I'm confident that I'll have a decent amount of data points by the time the presentation rolls around.
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