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Time-resolved fluorescence microscopy in high-resolution brain imaging

Published on June 30, 2024   30 min

A selection of talks on Methods

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0:00
Today, we'll talk about time-resolved fluorescence microscopy in high-resolution brain imaging.
0:10
This lecture will consist of two parts; the first one will be about fluorescence lifetime imaging, or FLIM, exemplified by the monitoring of intracellular nanomolar calcium concentrations, and the second part will be about time-resolved fluorescence anisotropy imaging which you'll try to explain by measuring molecular mobility on the nanoscale.
0:41
First, we have to understand why and when we actually need time-resolved imaging inflorescence microscopy of the brain. Well, you probably know that in the majority of cases, fluorescent indicators increase their emission intensity upon binding to the signal molecule of interest. However, the emission intensity signal might depend directly on the local concentration of the indicator as well as on the optical properties of the tissue. This actually makes it pretty difficult to assess quantitatively how much of the signal and molecule is reported by the fluorescence indicator, or historically, concentration measurements of signaling molecules were, therefore, achieved using so-called ratiometric indicators. The ratiometric indicators changed either the absorption or emission spectrum upon binding to the signaling molecule so that the ratio between different parts of the emission spectrum reports the signaling molecule concentration which, therefore, does not depend on the indicator concentration.

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Time-resolved fluorescence microscopy in high-resolution brain imaging

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