一個高效識別全腦神經(jīng)回路分子表型的成像系統(tǒng)
染色方法：
病毒示蹤標記
標記方法：
EGFP、EYFP、 ERFP、 mCherry、 Tdtomato
包埋方法：
樹脂包埋
成像平臺：
BioMapping 9000
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Figure 1. Phenotyping brain-wide neural circuits. (a) Presentation of the whole pipeline. (b) Schematic of the rapid whole-brain optical tomography system with automatic slice collection. (c) and (d) Typical coronal images of propidium iodide (PI)-stained 2 M C57BL/6 J mouse brains acquired by the system before and after optimization. Scale bar: 1 mm.
Figure 2. Automatic slice collection of imaging a Thy1-eYFP H-line mouse brain. (a) The brain was imaged by our system as 272 coronal sections at an interval of 50 μm, and all imaged tissue slices were sequentially collected during data acquisition. (b) All collected slices were placed on the slides and imaged with a Nikon Eclipse Ni-E wide-field microscope one-by-one manually. Blue and green represent 4′, 6-diamidino-2- phenylindole (DAPI) staining and YFP-labelled images of the collected slices, respectively. All images are arranged according to the corresponding in situ images, and the blank areas represent the missing slices. Enlarged views of the hippocampal coronal plane are indicated by white rectangles in row 9 and column 10. (c) An in situ image during whole-brain imaging with our system, and (d) the image of the corresponding collected slice imaged with a Zeiss LSM 710 confocal microscope. Scale bars in (c) and (d) are 1 mm.
Figure 3. Whole-brain mapping of inputs to MOp, SSp and VISp. (a) Schematic of direct inputs to targeted areas using stereotaxic injections of CAV-Cre. Left side shows the gene elements of the Ai14 Cre-reporter mouse and CAV-Cre used in a labelling experiment. Right side shows that CAV-Cre was injected into the targeted area of a mouse brain. (b) Whole-brain volume rendering of the inputs to MOp, SSp and VISp. (c,d) Quantification of the brain-wide inputs in cortical and subcortical brain areas to MOp, SSp and VISp. Error bars represent SEM. Others represent all other input regions. The significant differences between pairs are indicated by the p value (*p < 0.05, **p < 0.01, and ***p < 0.001). Injection sites were excluded from the data analysis.
Movie 1. Closeup of the system at a vertical view at runtime.
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Figure 1. Phenotyping brain-wide neural circuits. (a) Presentation of the whole pipeline. (b) Schematic of the rapid whole-brain optical tomography system with automatic slice collection. (c) and (d) Typical coronal images of propidium iodide (PI)-stained 2 M C57BL/6 J mouse brains acquired by the system before and after optimization. Scale bar: 1 mm.
Figure 2. Automatic slice collection of imaging a Thy1-eYFP H-line mouse brain. (a) The brain was imaged by our system as 272 coronal sections at an interval of 50 μm, and all imaged tissue slices were sequentially collected during data acquisition. (b) All collected slices were placed on the slides and imaged with a Nikon Eclipse Ni-E wide-field microscope one-by-one manually. Blue and green represent 4′, 6-diamidino-2- phenylindole (DAPI) staining and YFP-labelled images of the collected slices, respectively. All images are arranged according to the corresponding in situ images, and the blank areas represent the missing slices. Enlarged views of the hippocampal coronal plane are indicated by white rectangles in row 9 and column 10. (c) An in situ image during whole-brain imaging with our system, and (d) the image of the corresponding collected slice imaged with a Zeiss LSM 710 confocal microscope. Scale bars in (c) and (d) are 1 mm.
Figure 3. Whole-brain mapping of inputs to MOp, SSp and VISp. (a) Schematic of direct inputs to targeted areas using stereotaxic injections of CAV-Cre. Left side shows the gene elements of the Ai14 Cre-reporter mouse and CAV-Cre used in a labelling experiment. Right side shows that CAV-Cre was injected into the targeted area of a mouse brain. (b) Whole-brain volume rendering of the inputs to MOp, SSp and VISp. (c,d) Quantification of the brain-wide inputs in cortical and subcortical brain areas to MOp, SSp and VISp. Error bars represent SEM. Others represent all other input regions. The significant differences between pairs are indicated by the p value (*p < 0.05, **p < 0.01, and ***p < 0.001). Injection sites were excluded from the data analysis.
Movie 1. Closeup of the system at a vertical view at runtime.
2017年10月24日，華中科技大學武漢光電國家研究中心袁菁老師課題組，發(fā)展了一種快速的新型fMOST平臺，并設計了一種有效的方法來繪制同一個大腦的全腦結構和分子信息: 快速成像和切片以及自動采集所有切片; 通過快速數(shù)據(jù)瀏覽方便地選擇感興趣的切片，然后執(zhí)行選擇的切片的事后特定免疫染色。 這一平臺顯著提高了神經(jīng)電路分子表型分型的效率，并為特定電路的細胞類型分析提供了自動化和產業(yè)化的途徑。文章發(fā)表在《科學報道》雜志上。
參考文獻
參考文獻[1]:Jiang T, Long B, Gong H, Xu T, Li X, Duan Z, Li A, Deng L, Zhong Q, Peng X, Yuan J. A platform for efficient identification of molecular phenotypes of brain-wide neural circuits.Sci Rep. (2017) 24;7(1):13891.