Abstract:
Here, we provide experimental verification supporting the use of short-section imaging bundles for two-photon microscopy imaging of the mouse brain. The 8 mm long bundle is made of a pair of heavy-metal oxide glasses with a refractive index contrast of 0.38 to ensure a high numerical aperture NA = 1.15. The bundle is composed of 825 multimode cores, ordered in a hexagonal lattice with a pixel size of 14 μm and a total diameter of 914 μm. We demonstrate successful imaging through custom-made bundles with 14 μm resolution. As the input, we used a 910 nm Ti-sapphire laser with 140 fs pulse and a peak power of 9 × 104 W. The excitation beam and fluorescent image were transferred through the fiber imaging bundle. As test samples, we used 1 μm green fluorescent latex beads, ex vivo hippocampal neurons expressing green fluorescent protein and cortical neurons in vivo expressing the fluorescent reporter GCaMP6s or immediate early gene Fos fluorescent reporter. This system can be used for minimal-invasive in vivo imaging of the cerebral cortex, hippocampus, or deep brain areas as a part of a tabletop system or an implantable setup. It is a low-cost solution, easy to integrate and operate for high-throughput experiments.
摘要:
这里,我们提供了实验验证,支持使用短切片成像束对小鼠大脑进行双光子显微镜成像。8毫米长的束由一对折射率对比度为0.38的重金属氧化物玻璃制成,以确保高数值孔径NA=1.15。该套件由825个多模内核组成,以六边形晶格排列,像素大小为14μm,总直径为914μm。我们展示了通过具有14μm分辨率的定制束成功成像。作为输入,我们使用了910nm的钛蓝宝石激光器,脉冲为140fs,峰值功率为9×104W。激发光束和荧光图像通过光纤成像束传输。作为测试样本,我们使用了1μm绿色荧光乳胶珠,体外表达绿色荧光蛋白的海马神经元和体内表达荧光报告基因GCaMP6s或立即早期基因Fos荧光报告基因的皮质神经元。该系统可用于大脑皮层的微创体内成像,海马体,或大脑深处区域作为桌面系统或可植入装置的一部分。这是一个低成本的解决方案,易于集成和操作的高通量实验。