Dong Zhe, Feng Yu, Diego Keziah, Baggetta Austin M, Sweis Brian M, Pennington Zachary T, Lamsifer Sophia I, Zaki Yosif, Sangiuliano Federico, Philipsberg Paul A, Morales-Rodriguez Denisse, Kircher Daniel, Slesinger Paul, Shuman Tristan, Aharoni Daniel, Cai Denise J
Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Sci Adv. 2025 Jul 4;11(27):eadr6470. doi: 10.1126/sciadv.adr6470. Epub 2025 Jul 2.
Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals. To enable simultaneous acquisition of two fluorescent wavelengths, we incorporated two CMOS sensors into a single miniscope. To validate our dual-channel miniscope, we imaged hippocampal CA1 region that co-expressed a dynamic calcium indicator (GCaMP) and a static nuclear signal (dTomato) while mice ran on a linear track. Our results suggest that, even when neurons were registered across days using dTomato signals, hippocampal spatial coding changes over time. In conclusion, our dual-channel miniscope enables imaging of two fluorescence wavelengths with minimal cross-talk between the two channels, opening the doors to a multitude of previously inaccessible experimental possibilities.
小型化荧光显微镜（微型显微镜）能够对自由活动动物体内大量神经元的钙信号活动进行成像。传统上，微型显微镜只能记录单一荧光波长。在此，我们展示了一种开源双通道微型显微镜，它能够在自由活动的动物体内同时记录两个波长。为实现两个荧光波长的同时采集，我们将两个互补金属氧化物半导体（CMOS）传感器集成到一个微型显微镜中。为验证我们的双通道微型显微镜，我们在小鼠在直线轨道上奔跑时，对共表达动态钙指示剂（GCaMP）和静态核信号（dTomato）的海马CA1区进行了成像。我们的结果表明，即使使用dTomato信号在数天内对神经元进行配准，海马的空间编码也会随时间变化。总之，我们的双通道微型显微镜能够对两个荧光波长进行成像，且两个通道之间的串扰最小，为众多以前无法实现的实验可能性打开了大门。
