Fluorescence imaging of neural activity in cultured neural networks

Master/D opdracht (25 weken, 1000 uur)

(Onderdelen van deze opdracht kunnen ook als Bachelor opdracht worden uitgevoerd)

Background and problem statement

Changes in activity patterns of neuronal networks in vitro have been extensively studied in our lab by using the microelectrode array (MEA) recording system. However, little is known about the relation between calcium transients and electrical activity in these cultured neural networks.

Assignment

In this project we will specifically address the question how bursts in cultured neural networks emerge, and test the hypotheses that bursts start via pacemaker neurons or small networks of neurons.

Rat primary cortical cells will be cultured on a MEA and bulk-loaded with a fluorescent calcium probe that penetrates the cell membrane. One or two-photon excited fluorescence confocal microscopy will be used to measure changes in intracellular calcium concentrations due to neural activity in large groups of neurons. Another fluorescent probe will be used to specificly label astrocytes, which allows us to separate neuronal from glial calcium transients. We will compare the activity during spontaneous bursts and activity evoked by electrical stimulation of a small group of neurons. The dynamics of neuronal and glial calcium transients will be correlated with extracellularly recorded neural activity.

Based on the calcium imaging and electrical activity measurements a neuronal model for spontaneously bursting cultured networks will be constructed that takes into account intracellular calcium transients.

Components

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Literature study

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Experiments

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Analysis

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Thesis writing

Educational program

BSc/MSc

EL/BMT

Research theme

Neurotechnology and cellular engineering

Principal Investigator track

Joost le Feber, Richard van Wezel

Supervision and info

Joost le Feber