Antic Srdjan D, Yan Ping, Acker Corey D, Spagnola Olivia T, Erol Zehra Y, Baser Ozge, Loew Leslie M
Neuroscience, UConn Health, School of Medicine, Institute for Systems Genomics, Farmington, Connecticut, USA.
Institute for the Brain and Cognitive Sciences (IBACS), University of Connecticut, Storrs, Connecticut, USA.
J Biophotonics. 2025 Mar 18:e70008. doi: 10.1002/jbio.70008.
New voltage-sensitive ElectroFluor (EF) dyes that emit across the visible and near-infrared spectrum (e.g., 730 nm) were recently developed. We evaluated EF-530, EF-630, and EF-730p-dyes spectrally orthogonal to green fluorescent protein (GFP)-at excitation wavelengths outside the conventional 470 nm range used for GFP-based indicators. Although previously applied in cardiac voltage imaging, their performance in neuronal tissue remains untested. We performed side-by-side comparisons using population voltage imaging in mouse cerebral cortex slices at optimal excitation wavelengths (530, 630, and 730 nm) and assessed cross-channel signal bleed-through across four excitation wavelengths (475, 530, 630, and 730 nm). All dyes produced robust optical signals at their optimal wavelengths, though non-preferred channels exhibited bleed-through with distinct amplitudes, polarities, and photobleaching patterns. These results provide detailed quantifications of EF dye performance for neuronal population imaging.
最近开发出了新型的电压敏感型电荧光（EF）染料，其发射光谱覆盖可见光和近红外光谱范围（例如730nm）。我们评估了EF-530、EF-630和EF-730p染料，这些染料在与绿色荧光蛋白（GFP）光谱正交的情况下，于用于基于GFP的指示剂的传统470nm范围之外的激发波长下使用。尽管此前已应用于心脏电压成像，但其在神经元组织中的性能仍未得到测试。我们在小鼠大脑皮层切片中使用群体电压成像，在最佳激发波长（530、630和730nm）下进行了并排比较，并评估了在四个激发波长（475、530、630和730nm）下的跨通道信号渗漏情况。所有染料在其最佳波长下均产生了强大的光学信号，不过非首选通道表现出具有不同幅度、极性和光漂白模式的信号渗漏。这些结果为神经元群体成像的EF染料性能提供了详细的量化数据。
