ArticleCell calcium2012
A comparative assessment of fluo Ca2+ indicators in rat ventricular myocytes.
Article in Cell calcium, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
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- High-speed optical imaging with sCMOS pixel reassignment.Nature communications · 2024Article
- Zosuquidar: An Effective Molecule for Intracellular CaInternational journal of molecular sciences · 2024Article
- Synchronous force and CaCommunications biology · 2024Article
- Cellular shortening and calcium dynamics are improved by noisy stimulus in a model of cardiomyopathy.Scientific reports · 2023Article
- Antioxidant Activity ofMarine drugs · 2022Article
- Purinoreceptors and ectonucleotidases control ATP-induced calcium waveforms and calcium-dependent responses in microglia: Roles of P2 receptors and CD39 in ATP-stimulated microglia.Frontiers in physiology · 2022Article
- PIP2 depletion and altered endocytosis caused by expression of Alzheimer's disease-protective variant PLCγ2 R522.The EMBO journal · 2021Article
- Starvation-induced autophagy via calcium-dependent TFEB dephosphorylation is suppressed by Shigyakusan.PloS one · 2020Article
- Mitochondrial regulation of [CaCell cycle (Georgetown, Tex.) · 2018Article
- Osteocyte calcium signals encode strain magnitude and loading frequency in vivo.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- An optimized protocol for purification of functional islets of Langerhans.Laboratory investigation; a journal of technical methods and pathology · 2017Article
- Efficient Binding of the NOS1AP C-Terminus to the nNOS PDZ Pocket Requires the Concerted Action of the PDZ Ligand Motif, the Internal ExF Site and Structural Integrity of an Independent Element.Frontiers in molecular neuroscience · 2017Article
- Designing Molecular Probes To Prolong Intracellular Retention: Application to Nitroxide Spin Probes.Bioconjugate chemistry · 2016Article
- Multiple cytosolic calcium buffers in posterior pituitary nerve terminals.The Journal of general physiology · 2016Article
- A comparison of fluorescent Ca²⁺ indicators for imaging local Ca²⁺ signals in cultured cells.Cell calcium · 2015Article
- A novel, rapid method to quantify intraplatelet calcium dynamics by ratiometric flow cytometry.PloS one · 2015Article
- Article
- Calcium movement in cardiac mitochondria.Biophysical journal · 2014Article
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Authors and funding
4 authors.
Funding
Abstract
The fluo family of indicators is frequently used in studying Ca(2+) physiology; however, choosing which fluo indicator to use is not obvious. Indicator properties are typically determined in well-defined aqueous solutions. Inside cells, however, the properties can change markedly. We have characterized each of three fluo variants (fluo-2MA, fluo-3 and fluo-4) in two forms-the acetoxymethyl (AM) ester and the K(+) salt. We loaded indicators into rat ventricular myocytes and used confocal microscopy to monitor depolarization-induced fluorescence changes and fractional shortening. Myocytes loaded with the indicator AM esters showed significantly different Ca(2+) transients and fractional shortening kinetics. Loading the K(+) salts via whole-cell patch-pipette eliminated differences between fluo-3 and fluo-4, but not fluo-2MA. Cells loaded with different indicator AM esters showed different staining patterns-suggesting differential loading into organelles. Ca(2+) dissociation constants (K(d,Ca)), measured in protein-rich buffers mimicking the cytosol were significantly higher than values determined in simple buffers. This increase in K(d,Ca) (decrease in Ca(2+) affinity) was greatest for fluo-3 and fluo-4, and least for fluo-2MA. We conclude that the structurally-similar fluo variants differ with respect to cellular loading, subcellular compartmentalization, and intracellular Ca(2+) affinity. Therefore, judicious choice of fluo indicator and loading procedure is advisable when designing experiments.
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