Oct 15, 2019
Quantifying Biogenic Silica (bSi) Deposition Rates Adapted Method & Fluorescence Reading (PDMPO) via Fluorometer
- 1University of Tennessee, Knoxville;
- 2Miami University of Ohio;
- 3The University of Tennessee, Knoxville
- The Aquatic Microbial Ecology Research Group - AMERG (The Buchan, Zinser and Wilhelm labs)
- Great Lakes Center for Fresh Waters and Human Health
External link: http://wilhelmlab.utk.edu/
Collection Citation: Brittany N Zepernick, Matthew Saxton, Steven W Wilhelm 2019. Quantifying Biogenic Silica (bSi) Deposition Rates Adapted Method & Fluorescence Reading (PDMPO) via Fluorometer. protocols.io https://dx.doi.org/10.17504/protocols.io.735hqq6
Manuscript citation:
Brittany N. Zepernick1, Matthew A. Saxton2, Steven W. Wilhelm1 University of Tennessee Knoxville1, Miami University2 Adapted from: (Saxton et al, 2012), (LeBlanc and Hutchins 2005) Original Methods Leblanc, K., & Hutchins, D. A. (2005). New applications of a biogenic silica deposition fluorophore in the study of oceanic diatoms. Limnology and Oceanography: Methods, 3(10), 462-476. Saxton, M. A., D'souza, N. A., Bourbonniere, R. A., McKay, R. M. L., & Wilhelm, S. W. (2012). Seasonal Si: C ratios in Lake Erie diatoms—evidence of an active winter diatom community. Journal of Great Lakes Research, 38(2), 206-211.
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 10, 2019
Last Modified: October 15, 2019
Collection Integer ID: 28509
Abstract
This method can be used to assess and quantify the rate of silica deposition (bSi) over time in diatoms to determine their rate of frustule synthesis. This protocol has been adapted for the processing of both cultures as well as environmental samples when inoculated with PDMPO [2‐(4‐pyridyl)‐5‐((4‐(2dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole], which is a fluorescent dye that is co-deposited with silica during frustule biosynthesis in a 3230:1 Si:PDMPO (mol:mol) ratio.
Materials
MATERIALS
LysoSensor™ Yellow/Blue DND-160 - Special PackagingFisher ScientificCatalog #L7545
Utilize a TD-700 Laboratory Fluorometer linked to a computer monitor with the downloaded Turner software to obtain fluorescence readings of the samples
Safety warnings
See SDS (Safety Data Sheet) for hazards and safety warnings.
