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A Novel Nicotinamide Adenine Dinucleotide Correction Method for Intracellular Ca2+ Measurement with Fura-2-Analog in Live Cells
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Biologia
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JoVE Journal Biologia
A Novel Nicotinamide Adenine Dinucleotide Correction Method for Intracellular Ca2+ Measurement with Fura-2-Analog in Live Cells

A Novel Nicotinamide Adenine Dinucleotide Correction Method for Intracellular Ca2+ Measurement with Fura-2-Analog in Live Cells

DOI:
10.3791/59881-v

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05:58 min

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September 20, 2019

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, , ,
1Department of Physiology,University of Ulsan College of Medicine/Asan Medical Center, 2Asan Medical Center, 3Asan Medical Institute of Convergence Science and Technology

Capítulos

  • 00:04Título
  • 00:41Mitochondrial Fluoroprobe Loading
  • 01:22In Situ Isosbestic Fura-2-FF Point Identification
  • 02:40Background Signal Detection and Cell Area Correction
  • 03:30R Factor Measurement
  • 04:12Results: Representative Intracellular Ca2+ Measurement
  • 05:15Conclusion

Summary

Tadução automática

Tadução automática

Due to the spectral overlapping of the excitation and emission wavelengths of NADH and fura-2 analogs, the signal interference from both chemicals in live cells is unavoidable during quantitative measurement of [Ca2+]. Thus, a novel online correction method of NADH signal interference to measure [Ca2+] was developed.

Tags

Intracellular Ca2+ MeasurementFura-2-analogNADHIsobestic PointUV Excitable ProbesSignal ContaminationNADH Correction MethodLive Cell ImagingMyocytesSaponinCalcium-free SolutionMulti-parametric Measurement

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