Week 9+10: More Troubleshooting

The past two weeks I’ve been staining and looking at leaf samples under the microscope and I’ve been investigating my dry-ice glandular trichome extraction method.

According to literature, the abaxial (bottom) side of the leaf is high in trichomes, and I confirmed this by taking many different sized leaves and looking at them under the microscope.

After confirming that the leaves actually had glandular trichomes, I tried putting a glass slide into the middle of the falcon tube before vortexing the leaves to see if any of the glandular trichomes would fall onto the slide (which I could directly view under the microscope).

However, there was nothing–at most I could find few glandular trichomes. I thought that maybe the protocol didn’t work with mint leaves, since literature mostly focused on using plant flowers. However, my advisor reread the paper and pointed out that the researchers had been putting their plant tissue in liquid nitrogen before vortexing, which allowed the samples to get very cold (colder than -100C). I had been putting my samples in the -80C freezer which often was actually around -70C, so my advisor proposed that the freezer was not cold enough to properly freeze the plant tissue and allow me to fracture/shatter the glandular trichomes off.

We tested his idea on Saturday by submerging the plant tissue in liquid nitrogen, and after vortexing the sample for a very long time (which unfortunately destroyed a lot of the tissue), it seemed that a good deal of glandular trichomes had fallen off the leaves. I still need to confirm if this is actually the case by looking at the extracted samples, so hopefully all goes well!