The content describes a novel automated patch clamp technique called "patch-walking" that improves the efficiency of finding synaptic connections between neurons in brain tissue. The key highlights are:
Patch-walking involves using multiple patch clamp pipettes, where one pipette is cleaned and reused to obtain a new whole cell recording while maintaining the others. This allows for probing many more potential connections compared to the traditional approach of retracting all pipettes after each recording attempt.
Mathematical modeling shows that patch-walking can yield 80-92% more probed connections than the traditional method for experiments with 10-100 cells and 2-8 pipettes.
The authors built a dual-pipette automated patch clamp system and demonstrated the patch-walking approach. Out of 136 patch attempts, they achieved 71 successful whole cell recordings (52.2% success rate) and probed 29 paired recordings, finding 3 synaptic connections.
Patch-walking offers advantages such as less tissue damage, faster experiment time, and the ability to record from more cells before cell death, making it particularly useful for studying rare tissue samples like human brain.
The authors discuss future improvements to patch-walking, such as optimal pipette-cell assignment strategies to avoid collisions, and integrating techniques like channelrhodopsin-assisted circuit mapping.
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by Yip,M. C., G... 게시일 www.biorxiv.org 04-01-2024
https://www.biorxiv.org/content/10.1101/2024.03.30.587445v2더 깊은 질문