Use of nernstein voltage sensitive dyes in measuring transmembrane voltage
First Claim
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1. A method of generating a signal output which is sensitive to membrane potential, the method comprising:
- providing a first component comprising one or more membranes;
adding a cationic membrane permeable nucleic acid staining dye to the first component; and
, monitoring a first signal output from the cationic membrane permeable nucleic acid staining dye, wherein the first signal output is correlated with the transmembrane potential across the one or more membranes.
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Abstract
Transmembrane potential measurement methods using cationic dyes, and anionic dyes are provided. Compositions of the cationic and anionic dyes and microfluidic systems which include the dyes and membranes are provided in conjunction with processing elements for transmembrane potential measurements.
172 Citations
58 Claims
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1. A method of generating a signal output which is sensitive to membrane potential, the method comprising:
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providing a first component comprising one or more membranes;
adding a cationic membrane permeable nucleic acid staining dye to the first component; and
,monitoring a first signal output from the cationic membrane permeable nucleic acid staining dye, wherein the first signal output is correlated with the transmembrane potential across the one or more membranes. - View Dependent Claims (2, 3, 4, 5, 6, 14, 15, 16, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 58)
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7. A method of producing a signal which is dependent on transmembrane potential, the method comprising:
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flowing a first mixture comprising one or more membranes and at least two voltage sensitive membrane permeable dyes through a first channel region; and
,monitoring at least a first signal output from at least one of the at least two voltage sensitive membrane permeable dyes, thereby producing a signal which is dependent on the transmembrane potential across the one or more membranes. - View Dependent Claims (9, 10, 11, 12, 13, 17, 18, 19, 20, 21)
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8. A method of producing a signal which is dependent on transmembrane potential, the method comprising:
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flowing a first mixture comprising one or more membranes and one or more voltage sensitive dyes through a first channel region; and
,monitoring at least a first signal output from at least one of the voltage sensitive dyes, thereby producing a signal which is dependent on the transmembrane potential across the one or more membranes, wherein the voltage sensitive dyes comprise one or more membrane permeable redistributing dyes, which one or more membrane permeable dyes comprise one or more ionic dye wherein flow of the membrane permeable redistributing dyes across the membrane is detected by monitoring the one or more signal outputs from the membrane permeable redistributing dyes before an equilibrium distribution of the membrane permeable redistributing dyes is reached. - View Dependent Claims (57)
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36. A method of generating an optical signal which is sensitive to transmembrane potential, the method comprising:
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providing a first component comprising one or more membrane;
adding a cationic membrane permeable redistributing dye to the first component;
adding an anionic membrane permeable redistributing dye to the first component; and
,measuring a first signal output from the cationic dye and a second signal output from the anionic dye, wherein one or more of the first and second signal outputs comprises an optical signal output, thereby generating the optical signal which is sensitive to the transmembrane potential. - View Dependent Claims (37, 38, 39, 51)
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40. A method of generating an optical signal which is dependent on transmembrane potential, the method comprising:
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providing a first component comprising one or more membrane;
adding at least a first membrane permeable redistributing dye to the first component, wherein the first membrane permeable redistributing dye comprises an ion;
measuring one or more signal output from the first redistributing dye before an equilibrium dye distribution is established, which one or more signal output comprises at least one optical signal output, thereby providing the optical signal which is dependent on transmembrane potential. - View Dependent Claims (41, 42, 43, 44, 45, 46, 47, 48, 49, 50)
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52. A method of generating a signal output which is sensitive to membrane potential, the method comprising:
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providing a first component comprising one or more membranes;
adding a cationic membrane permeable nucleic acid staining dye to the first component, wherein the dye is Red-fluorescent dye SYTO 62; and
,monitoring a first signal output from the cationic membrane permeable nucleic acid staining dye, wherein the first signal output is correlated with the transmembrane potential across the one or more membranes.
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53. A method of producing a signal which is dependent on transmembrane potential, the method comprising:
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flowing a first mixture comprising one or more membranes and one or more voltage sensitive dyes through a first channel region, wherein the one or more voltage sensitive dyes comprise SYTO 62; and
,monitoring at least a first signal output from at least one of the voltage sensitive dyes, thereby producing a signal which is dependent on the transmembrane potential across the one or more membranes.
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54. A method of generating a signal output which is sensitive to membrane potential, the method comprising:
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providing a first component comprising one or more membranes;
adding a cationic membrane permeable nucleic acid staining dye to the first component;
adding an anionic membrane permeable redistributing dye to the first component;
monitoring a first signal output from the cationic membrane permeable nucleic acid staining dye, wherein the first signal output is correlated with the transmembrane potential across the one or more membranes; and
,measuring a second signal output from the anionic membrane permeable redistributing dye, thereby providing a further indication of changes in the transmembrane potential;
wherein the cationic membrane permeable nucleic acid staining dye is SYTO 62, at a concentration of between about 0.01 and about 50 μ
M and the anionic dye is DiBAC4(3), at a concentration of between about 0.01 and about 50 μ
M.
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55. A method of producing a signal which is dependent on transmembrane potential, the method comprising:
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flowing a first mixture comprising one or more membranes, one or more voltage sensitive dyes and an anionic membrane permeable redistributing dye through a first channel region;
monitoring at least a first signal output from at least one of the voltage sensitive dyes, thereby producing a signal which is dependent on the transmembrane potential across the one or more membranes; and
measuring a second signal output from the anionic membrane permeable redistributing dye, thereby providing a further indication of changes in the transmembrane potential;
wherein the cationic membrane permeable nucleic acid staining dye is SYTO 62, at a concentration of between about 0.01 and about 50 μ
M and the anionic dye is DiBAC4(3), at a concentration of between about 0.01 and about 50 μ
M.
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56. A method of generating an optical signal which is sensitive to transmembrane potential, the method comprising:
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providing a first component comprising one or more membrane;
adding a cationic membrane permeable redistributing dye comprising Syto 62 to the first component;
adding an anionic membrane permeable redistributing dye comprising DiBAC4(3) to the first component; and
,measuring a first signal output from the cationic dye and a second signal output from the anionic dye, wherein one or more of the first and second signal outputs comprises an optical signal output, thereby generating the optical signal which is sensitive to the transmembrane potential.
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Specification