Department of Electrical & Computer Engineering, Boston University, 8 Saint Mary's St., Boston, Massachusetts, 02215, USA.
Department of Biomedical Engineering, Boston University, 44 Cummington Mall, Boston, Massachusetts, 02215, USA.
Sci Rep. 2018 May 21;8(1):7921. doi: 10.1038/s41598-018-26240-8.
Fast volumetric microscopy is required to monitor large-scale neural ensembles with high spatio-temporal resolution. Widefield fluorescence microscopy can image large 2D fields of view at high resolution and speed while remaining simple and costeffective. A focal sweep add-on can further extend the capacity of widefield microscopy by enabling extended-depth-of-field (EDOF) imaging, but suffers from an inability to reject out-of-focus fluorescence background. Here, by using a digital micromirror device to target only in-focus sample features, we perform EDOF imaging with greatly enhanced contrast and signal-to-noise ratio, while reducing the light dosage delivered to the sample. Image quality is further improved by the application of a robust deconvolution algorithm. We demonstrate the advantages of our technique for in vivo calcium imaging in the mouse brain.
快速体积显微镜是监测具有高时空分辨率的大规模神经团的必要手段。宽场荧光显微镜可以以高分辨率和速度成像大的 2D 视场,同时保持简单和具有成本效益。焦点扫描附加组件可以通过实现扩展景深(EDOF)成像进一步扩展宽场显微镜的容量,但存在无法消除离焦荧光背景的问题。在这里,我们通过使用数字微镜设备仅针对聚焦的样本特征,在大大提高对比度和信噪比的同时,减少了施加到样本的光剂量,从而进行 EDOF 成像。通过应用强大的去卷积算法进一步提高了图像质量。我们展示了我们的技术在活体小鼠大脑钙成像中的优势。
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