Mechanism of the membrane potential sensitivity of the fluorescent membrane probe merocyanine 540

Dragsten, Paul R.; Webb, Watt W.

doi:10.1021/bi00617a024

Cited by 164 publications

(140 citation statements)

References 38 publications

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“…ANS binding to stimulated lymphocytes is markedly different from binding to unstimulated lymphocytes (1); it is unclear how much of the difference is due to differences in membrane potential between stimulated and unstimulated cells. Merocyanine 540 stains marrow cells but not peripheral blood lymphocytes (66); again, the change in the fluorescence of this dye in response to nerve action potentials, which results from changes in orientation and distribution of dye in the membrane (18), is minuscule (about one part in lo3), and one would not expect to be able to detect such changes in even the most precise of today's flow cytometers.…”

Section: Resultsmentioning

confidence: 99%

Flow cytometric probes of early events in cell activation

Shapiro

1981

Cytometry

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By now, it's well established that the surface of a cell Contains transmembrane proteins which, on binding ligands, tell The biochemical computer inside what to do Without a need for any of the ligands coming through. Receptor‐triggered cytoplasmic “interrupt routines” May lead the nucleus to activate new sets of genes, Or otherwise induce a cell, which never reasons why, To twitch, secrete, endocytose, or reproduce, or die. By merely watching labeled ligands bind, we cannot know Down which of many paths the cells thus tagged are apt to go, But other changes, shortly after binding, may predict Which of its several options any given cell has picked. Flow cytometric methods are described here which allow Membrane potential to be estimated, to show how Potential changes may occur in seconds after binding Of ligand to a functional receptor, or for finding No change when cells don't have the right receptor, or can't see The ligand for antagonists of like affinity. With chlorotetracycline fluorescence, one can find Release of membrane calcium soon after ligands bind; Both this and the potential measurement can help support Decisions in an instrument about which cells to sort, Though narrowing the distributions which these methods yield Would greatly help to broaden applications in this field.

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Section: Resultsmentioning

confidence: 99%

Flow cytometric probes of early events in cell activation

Shapiro

1981

Cytometry

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“…[13] VSDs have been useful optical indicators for membrane potential measurements in cells and organelles that are too small for electrodes. [14] VSDs are generally categorized by their response to the change in local electric field, and one distinguishes fast-response and slow-response probes. [15] Most slow-response VSDs exhibit polarity-dependent optical characteristics.…”

Section: Introductionmentioning

confidence: 99%

Potassium Sensitive Optical Nanosensors Containing Voltage Sensitive Dyes

2015

Chimia

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Ionophore-based ion-selective optical nanosensors have been explored for a number of years. Voltage sensitive dyes (VSDs) have been introduced into this type of sensors only very recently, forming a new class of analytical tools. Here, K + -sensitive nanospheres incorporating a lipophilic VSD were successfully fabricated and characterized. The nanosensors were readily delivered into the social amoeba Dictyostelium discoideum in a non-invasive manner, forming a promising new platform for intracellular ion quantification and imaging.

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“…[25] Here we compare the voltage sensitivity of the SHG from JR1 with that from three widely studied styryl dyes (FM4-64, di-4-ANEPPS, and RH237) in hemispherical lipid bilayers (HLBs). [15,26,27] The results show that the SHG signal from the porphyrin-based dye is exceptionally sensitive to an electric field.The sensitivity of the porphyrin dye JR1 and the styryl dyes FM4-64, di-4-ANEPPS, and RH237 to transmembrane potential was investigated using the setup shown in Figure 1. [15,26,27] Each dye was added to an aqueous solution of phosphate buffered saline to give a dye concentration of 5 mm.…”

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confidence: 99%

“…[15,26,27] The results show that the SHG signal from the porphyrin-based dye is exceptionally sensitive to an electric field.…”

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confidence: 99%

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Porphyrins for Probing Electrical Potential Across Lipid Bilayer Membranes by Second Harmonic Generation

et al. 2013

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Neurons communicate by using electrical signals, mediated by transient changes in the voltage across the plasma membrane. Optical techniques for visualizing these transmembrane potentials could revolutionize the field of neurobiology by allowing the spatial profile of electrical activity to be imaged in real time with high resolution, along individual neurons or groups of neurons within their native networks. [1,2] Second harmonic generation (SHG) is one of the most promising methods for imaging membrane potential, although so far this technique has only been demonstrated with a narrow range of dyes. [3] Here we show that SHG from a porphyrin-based membrane probe gives a fast electro-optic response to an electric field which is about 5-10 times greater than that of conventional styryl dyes. Our results indicate that porphyrin dyes are promising probes for imaging membrane potential.Studies of excitable cells, such as neurons and cardiac myocytes, require new methods for mapping changes in membrane potential, ideally with a voltage resolution of about 1 mV, a time resolution of 1 ms, and a spatial resolution of 1 mm. [1,2] Microelectrodes can be used to measure transmembrane potentials with excellent sensitivity and temporal resolution, but they do not provide subcellular spatial resolution. Fluorescent calcium indicators are widely used to probe membrane potential indirectly, through the intracellular Ca 2+ concentration. However, changes in calcium concentration do not accurately reflect voltage transients. [4] Fluorescent voltage-sensitive dyes were first developed 30 years ago, [5] and recent advances in the molecular design of these dyes [6][7][8][9][10] have made it possible to track the spatial evolution of action potentials. [11][12][13][14] Compared with fluorescence, SHG imaging has several advantages as a technique for probing membrane potential: [3,[15][16][17][18][19][20][21] SHG has a greater intrinsic sensitivity to electric fields than fluorescence, [19][20][21] and it is only produced by noncentrosymmetric molecules in noncentrosymmetric environments, thus making it ideal for probing interfaces such as lipid bilayers. Similar to twophoton-excited fluorescence, SHG is a two-photon nonlinear optical effect, and it brings the advantages of depth penetration associated with multiphoton microscopy. [22] The main disadvantage of SHG is that it often gives lower intensity than fluorescence; there is a need for the design of brighter, more voltage-sensitive SHG dyes. [3,23] Styryl dyes and retinal chromophores have been shown to exhibit voltage-sensitive SHG when localized in plasma membranes, [15,24] and the voltage sensitivity of their SHG is greater than that of their fluorescence. [19][20][21] Recently we reported that amphiphilic porphyrins such as JR1 exhibit strong SHG when localized in lipid membranes. [25] Here we compare the voltage sensitivity of the SHG from JR1 with that from three widely studied styryl dyes (FM4-64, di-4-ANEPPS, and RH237) in hemispherical lipid bilayers (HLBs). [15,26,27] ...

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Mechanism of the membrane potential sensitivity of the fluorescent membrane probe merocyanine 540

Cited by 164 publications

References 38 publications

Flow cytometric probes of early events in cell activation

Flow cytometric probes of early events in cell activation

Potassium Sensitive Optical Nanosensors Containing Voltage Sensitive Dyes

Porphyrins for Probing Electrical Potential Across Lipid Bilayer Membranes by Second Harmonic Generation

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