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. 1983 May;42(2):195–198. doi: 10.1016/S0006-3495(83)84386-0

Fluorescence monitoring of electrical responses from small neurons and their processes.

A Grinvald, A Fine, I C Farber, R Hildesheim
PMCID: PMC1329223  PMID: 6860775

Abstract

To improve the sensitivity of fluorescence measurements of electrical responses from small cells and their processes, we have optimized the optical measuring system. The fluorescence intensity from a stained cell was increased 40-fold relative to our previous apparatus. The increased fluorescence intensity permits the use of an inexpensive photodiode (or a photodiode array) that has a approximately 10-fold higher quantum efficiency relative to a photomultiplier. Utilizing the improved apparatus, we optically recorded an action potential of a 2 microns wide neuronal process with a signal-to-noise ratio of approximately 50 (root mean square noise) without averaging. We also report the design of an improved fluorescence voltage-sensitive probe; the fractional change of the fluorescence signal under optimal conditions was 21%/100 mV.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Calvet J., Calvet M. C. A simple device for making a standard inverted phase-contrast microscope movable. J Neurosci Methods. 1981 Aug;4(2):105–108. doi: 10.1016/0165-0270(81)90043-1. [DOI] [PubMed] [Google Scholar]
  2. Cohen L. B., Salzberg B. M., Davila H. V., Ross W. N., Landowne D., Waggoner A. S., Wang C. H. Changes in axon fluorescence during activity: molecular probes of membrane potential. J Membr Biol. 1974;19(1):1–36. doi: 10.1007/BF01869968. [DOI] [PubMed] [Google Scholar]
  3. Cohen L. B., Salzberg B. M., Grinvald A. Optical methods for monitoring neuron activity. Annu Rev Neurosci. 1978;1:171–182. doi: 10.1146/annurev.ne.01.030178.001131. [DOI] [PubMed] [Google Scholar]
  4. Cohen L. B., Salzberg B. M. Optical measurement of membrane potential. Rev Physiol Biochem Pharmacol. 1978;83:35–88. doi: 10.1007/3-540-08907-1_2. [DOI] [PubMed] [Google Scholar]
  5. Giloh H., Sedat J. W. Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. Science. 1982 Sep 24;217(4566):1252–1255. doi: 10.1126/science.7112126. [DOI] [PubMed] [Google Scholar]
  6. Grinvald A., Cohen L. B., Lesher S., Boyle M. B. Simultaneous optical monitoring of activity of many neurons in invertebrate ganglia using a 124-element photodiode array. J Neurophysiol. 1981 May;45(5):829–840. doi: 10.1152/jn.1981.45.5.829. [DOI] [PubMed] [Google Scholar]
  7. Grinvald A., Farber I. C. Optical recording of calcium action potentials from growth cones of cultured neurons with a laser microbeam. Science. 1981 Jun 5;212(4499):1164–1167. doi: 10.1126/science.7233210. [DOI] [PubMed] [Google Scholar]
  8. Grinvald A., Hildesheim R., Farber I. C., Anglister L. Improved fluorescent probes for the measurement of rapid changes in membrane potential. Biophys J. 1982 Sep;39(3):301–308. doi: 10.1016/S0006-3495(82)84520-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grinvald A., Ross W. N., Farber I. Simultaneous optical measurements of electrical activity from multiple sites on processes of cultured neurons. Proc Natl Acad Sci U S A. 1981 May;78(5):3245–3249. doi: 10.1073/pnas.78.5.3245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gupta R. K., Salzberg B. M., Grinvald A., Cohen L. B., Kamino K., Lesher S., Boyle M. B., Waggoner A. S., Wang C. H. Improvements in optical methods for measuring rapid changes in membrane potential. J Membr Biol. 1981 Feb 15;58(2):123–137. doi: 10.1007/BF01870975. [DOI] [PubMed] [Google Scholar]
  11. Kimhi Y., Palfrey C., Spector I., Barak Y., Littauer U. Z. Maturation of neuroblastoma cells in the presence of dimethylsulfoxide. Proc Natl Acad Sci U S A. 1976 Feb;73(2):462–466. doi: 10.1073/pnas.73.2.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ross W. N., Salzberg B. M., Cohen L. B., Grinvald A., Davila H. V., Waggoner A. S., Wang C. H. Changes in absorption, fluorescence, dichroism, and Birefringence in stained giant axons: : optical measurement of membrane potential. J Membr Biol. 1977 May 6;33(1-2):141–183. doi: 10.1007/BF01869514. [DOI] [PubMed] [Google Scholar]
  13. Waggoner A. S. Dye indicators of membrane potential. Annu Rev Biophys Bioeng. 1979;8:47–68. doi: 10.1146/annurev.bb.08.060179.000403. [DOI] [PubMed] [Google Scholar]

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