Geochemistry & Ecology of Red Mat Systems (GERMS)

Undergraduate Summer Research Program

Week Three

Microscopy, Enumeration, and Digital Imaging:

Mat Layer Processing:

Site: Imperial

Image – Visible Light

10X	40X

Image – F Fluorescence

10X	40X

Image – R Fluorescence

10X	40X

Grid-Based Counts:

Grid @ 10X	Grid # 40 X

Chemical Analysis of Acid-Digested Samples for Metal Assessment

Specific Post-Acid Digestion Results:

Aluminum	0.007 mg/L
Copper	0.00 mg/L
Total Iron	0.06 mg/L
Zinc	0.52 mg/L
Manganese	Not preformed

Pigment Analysis

Methanol Extraction:

Methanol Extraction Procedure: We first spun test tubes for 5 minutes in a centrifuge to bring down the cells to the bottom of the test tube. We then added methanol and re-suspended the cell. We then wrapped the test tube in foil and left it sit on ice for 30 minutes to let the methanol extract the pigments.
Methanol Extraction Results:

In vivo Bead-Beating:

In vivo Bead-Beating Procedure: e filled a tube halfway with.1mm zirconium beads, added GTE, and our sample. We then placed that tube into the bead beater. After bead beating the tube we spun it for minutes in a centrifuge and placed the tube on ice.
In vivo Bead-Beating Procedure:

Discussion:

This week we were analyzing the data we collected. Liz and I started off by doing out pigment analysis. We then moved onto doing post-acid digestion test on the water that we collected while in Yellowstone. The last day of the week Liz and I spent working on the microscope, counting the filaments from our layers. I was able to count only 3 within the grid, which was unfortunate. The pictures turned out great from the microscope and I had an amazing time working with fluorescence for detecting pigments and various bacteria within the mats.