Fast Optical Sectioning for Widefield Fluorescence Mesoscopy with the Mesolens based on HiLo Microscopy.
Fast Optical Sectioning for Widefield Fluorescence Mesoscopy with the Mesolens based on HiLo Microscopy.
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DOI：
10.1038/s41598-018-34516-2
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发表时间：
2018-11-02
影响因子：
4.6
通讯作者：
McConnell G
中科院分区：
文献类型：
--
作者：
Schniete J;Franssen A;Dempster J;Bushell TJ;Amos WB;McConnell G
我们在这里提出了一种基于HiLo显微镜的快速光学切片方法，该方法可以在17小时内对体积高达4.4 mm × 3 mm × 3 mm的标本进行成像（估计包括1000张图像的z堆栈，不包括计算时间），整个过程具有亚细胞分辨率。宽场落射荧光成像是使用Mesolens进行的，使用高像素数的相机，能够传感器移位，以生成259.5百万像素的图像，我们已经开发了定制软件来执行非常大的数据集的HiLo处理。使用这种方法，我们获得了与共聚焦激光扫描显微镜（CLSM）相当的切片强度，切片薄至6.8 ± 0.2 μm，原始采集速度为1分钟/切片，在4.4 mm的Mesolens的全视场（FOV）上比CLSM快30倍，横向分辨率为0.7 μm，轴向分辨率为7 μm。我们已经应用这种HiLo介观方法图像固定和荧光染色的海马神经元标本和5天大的斑马鱼幼虫。
We present here a fast optical sectioning method for mesoscopy based on HiLo microscopy, which makes possible imaging of specimens of up to 4.4 mm × 3 mm × 3 mm in volume in under 17 hours (estimated for a z-stack comprising 1000 images excluding computation time) with subcellular resolution throughout. Widefield epifluorescence imaging is performed with the Mesolens using a high pixel-number camera capable of sensor-shifting to generate a 259.5 Megapixel image, and we have developed custom software to perform HiLo processing of the very large datasets. Using this method, we obtain comparable sectioning strength to confocal laser scanning microscopy (CLSM), with sections as thin as 6.8 ± 0.2 μm and raw acquisition speed of 1 minute per slice which is up to 30 times faster than CLSM on the full field of view (FOV) of the Mesolens of 4.4 mm with lateral resolution of 0.7 μm and axial resolution of 7 μm. We have applied this HiLo mesoscopy method to image fixed and fluorescently stained hippocampal neuronal specimens and a 5-day old zebrafish larva.
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影响因子：
3.4
作者：
Dean, Kevin M.;Roudot, Philippe;Welf, Erik S.;Danuser, Gaudenz;Fiolka, Reto
通讯作者：
Fiolka, Reto
DOI：
10.1016/b978-0-12-374920-8.00203-4
发表时间：
2011-01-01
期刊：
COMPREHENSIVE BIOPHYSICS, VOL 2: BIOPHYSICAL TECHNIQUES FOR CHARACTERIZATION OF CELLS
影响因子：
--
作者：
Amos, B.;McConnell, G.;Wilson, T.
通讯作者：
Wilson, T.
影响因子：
3.4
作者：
McNair, K;Davies, CH;Cobb, SR
通讯作者：
Cobb, SR
影响因子：
48
作者：
Vettenburg, Tom;Dalgarno, Heather I. C.;Dholakia, Kishan
通讯作者：
Dholakia, Kishan
影响因子：
48
作者：
Tokunaga, Makio;Imamoto, Naoko;Sakata-Sogawa, Kumiko
通讯作者：
Sakata-Sogawa, Kumiko