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Patch-walking, a novel automated multi-pipette approach, significantly improves the efficiency of probing synaptic connections between neurons in brain tissue compared to traditional methods.


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Significant technical challenges exist when measuring synaptic connections between neurons in living brain tissue. The patch clamping technique, when used to probe for synaptic connections, is manually laborious and time-consuming. To improve its efficiency, we pursued another approach: instead of retracting all patch clamping electrodes after each recording attempt, we cleaned just one of them and reused it to obtain another recording while maintaining the others. With one new patch clamp recording attempt, many new connections can be probed. By placing one pipette in front of the others in this way, one can “walk” across the tissue, termed “patch-walking.” We performed 136 patch clamp attempts for two pipettes, achieving 71 successful whole cell recordings (52.2%). Of these, we probed 29 pairs (i.e., 58 bidirectional probed connections) averaging 91 µ m intersomatic distance, finding 3 connections. Patch-walking yields 80-92% more probed connections, for experiments with 10-100 cells than the traditional synaptic connection searching method.

Motivation Recognizing the manual labor and time-intensive nature of patch clamping when trying to find synaptic connections, we aim to improve its efficiency. We introduce a novel approach, termed “patch-walking,” where one patch clamping electrode is cleaned and reused, enabling the exploration of numerous connections with a single recording attempt and improving the efficiency of identifying synaptic connections.

### Competing Interest Statement

I.K., W.M.S, and C.R.F. are co-inventors on a patent describing pipette cleaning that is licensed by Sensapex.

Patch-walking: Coordinated multi-pipette patch clamp for efficiently finding synaptic connections

automated-patch-walking-an-efficient-approach-for-probing-synaptic-connections-in-brain-tissue


Automated Patch-Walking: An Efficient Approach for Probing Synaptic Connections in Brain Tissue


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Automated Multi-Pipette Patch Clamp for Synaptic Connectivity Mapping