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

janelia7_blocks-janelia7_biblio_header | block

eLife. 2024 Nov 18;13:RP97399. doi: 10.7554/elife.97399

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

Yip Mighten, Gonzalez Mercedes, Lewallen Colby, Landry Corey, Kolb Ilya, Yang Bo, Stoy William, Fong Ming-fai, Rowan Matthew, Boyden Edward, Forest Craig

janelia7_blocks-janelia7_biblio_abstract | block

Abstract

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 mouse brain slice, 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 μm intersomatic distance, finding three connections. Patch-walking yields 80-92% more probed connections, for experiments with 10-100 cells than the traditional synaptic connection searching method.

Main Menu (Mobile)- Block

Main Menu - Block

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

Abstract

Janelia Authors