Related Experiment Video
Updated: Jun 20, 2026

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery
Published on: October 4, 2018
Detecting the phosphate status of phytoplankton by enzyme-labelled fluorescence and flow cytometry
Marco Dignum1, Hans L Hoogveld, Hans C P Matthijs
1Centre for Limnology, Netherlands Institute of Ecology, Royal Netherlands Academy of Arts and Sciences, Nieuwersluis, The Netherlands. m.dignum@nioo.knaw.nl
Abstract:
The novel phosphatase substrate, ELF-97 phosphate, yields intensely green fluorescent precipitates of ELF-97 alcohol (ELFA) upon enzymatic dephosphorylation, and thereby traces phosphatase activity back to its producer. In this study, we show that ELFA fluorescence is a useful tool in flow cytometric analysis of natural phytoplankton populations. Presence of endogenous fluorescent pigments allowed flow cytometric distinction of clusters in the phytoplankton community in Lake Loosdrecht (The Netherlands): Eukaryotes (diatoms and green algae), chlorophyll a and b containing but phycobilin-less cyanobacteria (Prochlorothrix hollandica), and phycocyanin-containing cyanobacteria (predominantly Limnothrix sp.). Several, but not all tested cyanobacteria showed ELFA fluorescence. The dominant Limnothrix sp. possesses a derepressible phosphatase, whereas the second most abundant strain, P. hollandica, did not have phosphatase activity. Within both natural and cultured populations of Limnothrix sp. we found discernible levels of ELFA fluorescence, indicating the presence of subpopulations with different physiological characteristics.
Related Concept Videos
Fluorescence and Phosphorescence: Instrumentation
Protein Dynamics in Living Cells
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Photoluminescence: Applications
Labeling DNA Probes
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

