@article {69325, title = {Patch-walking: Coordinated multi-pipette patch clamp for efficiently finding synaptic connections}, journal = {eLife}, volume = {13}, year = {2024}, month = {2024 Nov 18}, pages = {RP97399}, 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 {\textquoteright}walk{\textquoteright} across the mouse brain slice, termed {\textquoteright}patch-walking.{\textquoteright} 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.

}, doi = {10.7554/elife.97399}, url = {https://doi.org/10.7554/elife.97399}, author = {Mighten C. Yip and Mercedes M. Gonzalez and Colby F. Lewallen and Corey R. Landry and Ilya Kolb and Bo Yang and William M. Stoy and Ming-fai Fong and Matthew JM Rowan and Edward S. Boyden and Craig R. Forest} } @article {67849, title = {Rapid reconstruction of neural circuits using tissue expansion and lattice light sheet microscopy}, journal = {eLife}, year = {2022}, month = {2022 Oct 26}, abstract = {

Electron microscopy (EM) allows for the reconstruction of dense neuronal connectomes but suffers from low throughput, limiting its application to small numbers of reference specimens. We developed a protocol and analysis pipeline using tissue expansion and lattice light-sheet microscopy (ExLLSM) to rapidly reconstruct selected circuits across many samples with single synapse resolution and molecular contrast. We validate this approach in\ Drosophila, demonstrating that it yields synaptic counts similar to those obtained by EM, can be used to compare counts across sex and experience, and to correlate structural connectivity with functional connectivity. This approach fills a critical methodological gap in studying variability in the structure and function of neural circuits across individuals within and between species.

}, doi = {10.7554/eLife.81248}, url = {https://doi.org/10.7554/eLife.81248}, author = {Joshua L. Lillvis and Hideo Otsuna and Xiaoyu Ding and Igor Pisarev and Takashi Kawase and Jennifer Colonell and Konrad Rokicki and Cristian Goina and Ruixuan Gao and Amy Hu and Kaiyu Wang and John Bogovic and Daniel E. Milkie and Edward S. Boyden and Stephan Saalfeld and Paul W. Tillberg and Barry J. Dickson} }