Synapse Segmentation Using Pixel-based Analysis of iGluSnFR3 Fluorescence Intensity

The Intensity-based Glutamate-Sensing Fluorescent Reporter (iGluSnFR) systemis a sensitive imaging tool that offers spatially and temporally resolved detection of excitatory neurotransmitter release events. We employed the recently developed iGluSnFR3 probe to study synaptic activity in hIPSC-derived neurons.

In our analyses of single-plane images, we found that the axonal trafficking of the probe was misidentified as synapses by current tools, and the detection of release events was inaccurate. To overcome this, we developed an image processing pipeline that employs stringent pixel selection and pixel clustering approaches to detect spatial and temporal peaks from synaptic activities.

Poster presented as part of the Crick BioImage Analysis Symposium 2024.

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