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Two-Photon Microscopy of Single Molecules

Published online by Cambridge University Press:  02 July 2020

R. Yuste ,
A. Majewska ,
K. Holthoff  and
K. Holthoff
R. Yuste
Affiliation:
Dept. of Biological Sciences, Columbia University, New York, NY, 10027
A. Majewska
Affiliation:
Dept. of Biological Sciences, Columbia University, New York, NY, 10027
K. Holthoff
Affiliation:
Dept. of Biological Sciences, Columbia University, New York, NY, 10027
K. Holthoff
Affiliation:
Dept. of Biological Sciences, Columbia University, New York, NY, 10027
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Extract

Two-photon excitation has enabled investigators to image living cells in highly scattering media like the central nervous system (1). We have used a custom-built two-photon microscope to image dendritic spines from living cortical pyramidal neurons. Pyramidal cells form the majority of the neuron in the mammalian cortex and they receive practically all their synaptic contacts through dendritic spines. Dendritic spines are small (<1 μm diameter) appendages that have been practically inaccessible to physiological measurements until the application of two-photon excitation to their study (2). We have concentrated in two questions:

A- Calcium compartmentalization of spines: Mechanisms of calcium decay kinetics.

Dendritic spines can compartmentalize calcium (2). Although the mechanisms of calcium influx into spines have been explored (3), it is unknown what determines the calcium decay kinetics in spines. We investigate calcium dynamics in spines from rat CA1 pyramidal neurons in slices.

Type
Advances in Multi-Photon imaging
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 804 - 805
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Denk, et al.(1994) J. Neurosci. Meth. 54: 151–162.CrossRefGoogle Scholar
2.Yuste, and Denk, . (1995). Nature 375: 682–684.CrossRefGoogle Scholar
3.Yuste, et al. (1999), J. Neurosci. 19 (6): 1976–1987.CrossRefGoogle Scholar
4.Majewska, et al. (2000). J. Neurosci. 20 (5), 1–13.Google Scholar
5.Dunaevsky, et al. (1999). Proc. Natl. Acad. Sci. USA 96 (23): 13438–13443.Google Scholar