利用淬滅熒光蛋白分子的化學(xué)再活化實(shí)現(xiàn)樹(shù)脂嵌入式熒光顯微成像
染色方法：
轉(zhuǎn)基因熒光標(biāo)記
標(biāo)記方法：
GFP、 YFP、 RFP
包埋方法：
瓊脂糖包埋
成像平臺(tái)：
BioMapping 5000
Figure 4 | Imaging of the thy1-YFPH mouse brain. (a) 3D presentation of a P21 thy1-YFPH mouse brain data set. Images were acquired with a voxel size of 0.5?0.5?1mm 3 . The inset image shows the contours of the whole mouse brain, reconstructed by autofluorescence and coronal plane sectioning on the blue labelled region (centred at the bregma þ1.50 level). Structures of the cerebral cortex are highlighted (white boxing inset) and enlarged to show details. The z axis represents the buffer penetration direction. (b) A layer Vpyramidal neuron recorded at a 0.2?0.2?0.2mm 3 voxel size on a commercial confocal microscope (Zeiss, LSM710). c–e show enlargements of the fine structures from the white box region of b. White arrows and numerals indicate (i) stubby, (ii) branched, (iii) thin, (iv) filopodium and (v) mushroom spines, as well as (vi) terminaux boutons, (vii) en passant bouton and (viii) structures suspected as synaptic connections. Scale bar: (a) inset, 1mm; (b), 20mm; (c,d), 2mm. Dimensions: (a) 1,850x620x1,000mm 3 ; (e) 9x18x9mm 3 .
Movie 1: Traveling in the brain
Figure 4 | Imaging of the thy1-YFPH mouse brain. (a) 3D presentation of a P21 thy1-YFPH mouse brain data set. Images were acquired with a voxel size of 0.5?0.5?1mm 3 . The inset image shows the contours of the whole mouse brain, reconstructed by autofluorescence and coronal plane sectioning on the blue labelled region (centred at the bregma þ1.50 level). Structures of the cerebral cortex are highlighted (white boxing inset) and enlarged to show details. The z axis represents the buffer penetration direction. (b) A layer Vpyramidal neuron recorded at a 0.2?0.2?0.2mm 3 voxel size on a commercial confocal microscope (Zeiss, LSM710). c–e show enlargements of the fine structures from the white box region of b. White arrows and numerals indicate (i) stubby, (ii) branched, (iii) thin, (iv) filopodium and (v) mushroom spines, as well as (vi) terminaux boutons, (vii) en passant bouton and (viii) structures suspected as synaptic connections. Scale bar: (a) inset, 1mm; (b), 20mm; (c,d), 2mm. Dimensions: (a) 1,850x620x1,000mm 3 ; (e) 9x18x9mm 3 .
Movie 1: Traveling in the brain
2014年6月2日，華中科技大學(xué)武漢光電國(guó)家研究中心駱清銘教授課題組發(fā)現(xiàn)綠色熒光蛋白在樹(shù)脂包埋的過(guò)程中熒光重激活方法，使用fMOST技術(shù)，研究小組演示了如何獲取密集的全腦神經(jīng)網(wǎng)絡(luò)數(shù)據(jù)。文章發(fā)表在《自然-通訊》雜志。
參考文獻(xiàn)
參考文獻(xiàn)[1]:Xiong H, Zhou Z, Zhu M, Lv X, Li A, Li S, Li L, Yang T, Wang S, Yang Z, Xu T, Luo Q, Gong H, Zeng S. Chemical reactivation of quenched fluorescent protein molecules enables resin-embedded fluorescence microimaging. Nat Commun. (2014);5:3992.
武漢東湖新技術(shù)開(kāi)發(fā)區(qū)豹澥街道高科園三路9號(hào)武漢光谷精準(zhǔn)醫(yī)療產(chǎn)業(yè)基地2.1期13棟12層