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Titolo:
Two-photon imaging in living brain slices
Autore:
Mainen, ZF; Maletic-Savatic, M; Shi, SH; Hayashi, Y; Malinow, R; Svoboda, K;
Indirizzi:
Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA Cold Spring Harbor Lab Cold Spring Harbor NY USA 11724 rbor, NY 11724 USA
Titolo Testata:
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
fascicolo: 2, volume: 18, anno: 1999,
pagine: 231 -
SICI:
1046-2023(199906)18:2<231:TIILBS>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEINS; LONG-TERM POTENTIATION; DENDRITIC SPINES; CALCIUM DYNAMICS; MICROSCOPY; NEURONS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
31
Recensione:
Indirizzi per estratti:
Indirizzo: Svoboda, K Cold Spring Harbor Lab, 1 Bungtown Rd, Cold Spring Harbor, NY 11724 USA Cold Spring Harbor Lab 1 Bungtown Rd Cold Spring Harbor NY USA 11724
Citazione:
Z.F. Mainen et al., "Two-photon imaging in living brain slices", METHODS, 18(2), 1999, pp. 231

Abstract

Two-photon excitation laser scanning microscopy (TPLSM) has become the tool of choice for high-resolution fluorescence imaging in intact neural tissues. Compared with other optical techniques, TPLSM allows high-resolution imaging and efficient detection of fluorescence signal with minimal photobleaching and phototoxicity. The advantages of TPLSM are especially pronounced in highly scattering environments such as the brain slice. Here we describeour approaches to imaging various aspects of synaptic function in living brain slices. To combine several imaging modes together with patch-clamp electrophysiological recordings we found it advantageous to custom-build an upright microscope. Our design goals were primarily experimental convenience and efficient collection of fluorescence. We describe our TPLSM imaging system and its performance in detail. We present dynamic measurements of neuronal morphology of neurons expressing green fluorescent protein (GFP) and GFP fusion proteins as well as functional imaging of calcium dynamics in individual dendritic spines. Although our microscope is a custom instrument, its hey advantages can be easily implemented as a modification of commercial laser scanning microscopes. (C) 1999 Academic Press.

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Documento generato il 05/12/20 alle ore 01:24:26