Practical Aspects of Measuring Intracellular Calcium Signals with Fluorescent Indicators

Kao, Joseph P. Y.; Gong, Li; Auston, Darryl A.

doi:10.1016/b978-0-12-374841-6.00005-0

Cited by 52 publications

(44 citation statements)

References 19 publications

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“…This level of intracellular loading is comparable to what has been achieved when cells are incubated with the AM esters of common fluorescent polycarboxylate ion indicators, 16–18 but is much lower than what we have achieved by incubation with the AM esters of nitroxides 1 , 2 , and 3 . 1, 19 The simplest rationale for the difference in loading is the aqueous solubility of the AM esters.…”

Section: Resultssupporting

confidence: 81%

Designing Molecular Probes To Prolong Intracellular Retention: Application to Nitroxide Spin Probes

et al. 2016

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Targeted delivery of molecular probes into cells enables cellular imaging through optical and magnetic modalities. Probe molecules that are well retained by cells can accumulate to higher intracellular concentrations, and thus increase the signal-to-noise ratio of, and widen the temporal window for, imaging. Here we synthesize a paramagnetic spin probe bearing six ionic functional groups and show that it has long intracellular half-life (> 12 hours) and exceptional biostability in living cells. We demonstrate that judicious incorporation of ionic substituents on probe molecules systematically increases intracellular retention time, and should therefore be beneficial to imaging experiments.

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Section: Resultssupporting

confidence: 81%

Designing Molecular Probes To Prolong Intracellular Retention: Application to Nitroxide Spin Probes

et al. 2016

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“…Background fluorescence was recorded after end-of-experiment cell lysis with 40 M digitonin (Sigma). At least 100 cells were analyzed from each imaged field; a region-ofinterest was defined for each cell, and the mean intensity within each region-of-interest was determined for each image frame and analyzed as described previously (37,38).…”

Section: Methodsmentioning

confidence: 99%

Proteolytic Activation of the Protease-activated Receptor (PAR)-2 by the Glycosylphosphatidylinositol-anchored Serine Protease Testisin

Driesbaugh

Buzza

Martin

et al. 2015

Journal of Biological Chemistry

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“…KmTx 2 (1 μg/ml stock solution in DMSO) was directly bath-applied with gentle mixing. Data reduction and analysis, including the calibration of microfluorimetric measurments to obtain [Ca 2+ ] i or [Na+] i , were performed as previously described (for Ca2+: Kao, 1994; Kao et al 2010; for Na + : Harootunian et al 1989). Origin software (OriginLab Corp., Northampton, MA) was used for data reduction and analysis.…”

Section: Methodsmentioning

confidence: 99%

The cytotoxic mechanism of karlotoxin 2 (KmTx 2) from Karlodinium veneficum (Dinophyceae)

et al. 2015

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This study demonstrates that the polyketide toxin karlotoxin 2 (KmTx 2) produced by Karlodinium veneficum, a dinoflagellate associated with fish kills in temperate estuaries worldwide, alters vertebrate cell membrane permeability. Microfluorimetric and electrophysiological measurements were used to determine that vertebrate cellular toxicity occurs through non-selective permeabilization of plasma membranes, leading to osmotic cell lysis. Previous studies showed that KmTx 2 is lethal to fish at naturally-occurring concentrations measured during fish kills, while sub-lethal doses severely damage gill epithelia. This study provides a mechanistic explanation for the association between K. veneficum blooms and fish kills that has long been observed in temperate estuaries worldwide.

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Practical Aspects of Measuring Intracellular Calcium Signals with Fluorescent Indicators

Cited by 52 publications

References 19 publications

Designing Molecular Probes To Prolong Intracellular Retention: Application to Nitroxide Spin Probes

Designing Molecular Probes To Prolong Intracellular Retention: Application to Nitroxide Spin Probes

Proteolytic Activation of the Protease-activated Receptor (PAR)-2 by the Glycosylphosphatidylinositol-anchored Serine Protease Testisin

The cytotoxic mechanism of karlotoxin 2 (KmTx 2) from Karlodinium veneficum (Dinophyceae)

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