Quantifying diatom silicification with the fluorescent dye, PDMPO

Limnology and Oceanography, Methods
Heather M McNairJeffrey W Krause

Abstract

Diatoms require silicic acid to construct ornately detailed cell walls called frustules. The growth and geographic distribution of diatoms is often controlled by the availability of silicic acid. Analytical methods exist to assess diatom community biogenic silica (bSiO2) production, but partitioning production among taxa has been largely qualitative. We present a method for the quantitative analysis of taxa-specific silica production through labeling diatoms with the fluorescent dye PDMPO [2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole]. To make PDMPO a quantitative tool: diatom frustules were solubilized to assess the total diatom community incorporation by quantitation of PDMPO fluorescence using a fluorometer, and laser confocal microscopy was used to quantify the fluorescence of PDMPO in single diatom cells. We created a fluorescence standard to intercalibrate the raw fluorescence signals of the fluorometer and microscope and to determine the fluorescence per mole of PDMPO. PDMPO incorporation was converted to silica production using diatom bSiO2:PDMPO incorporation ratios which varied systematically with silicic acid concentration. Above 3 μM Si(OH)4, bSiO2:PDMPO was constant and PDMPO incor...Continue Reading

References

Oct 28, 1998·Proceedings of the National Academy of Sciences of the United States of America·R E TurnerJ Cope
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Nov 20, 2013·Proceedings of the National Academy of Sciences of the United States of America·Philipp AssmyDieter Wolf-Gladrow

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Citations

Jun 26, 2018·Limnology and Oceanography·Heather M McNairJeffrey W Krause
Oct 30, 2020·Environmental Microbiology·Penelope A AjaniShauna A Murray
Oct 26, 2021·Frontiers in Microbiology·Phoebe A ArgyleMartina A Doblin

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