塑性包埋大體積免疫標(biāo)記樣品的三維高分辨率成像
染色方法：
整體免疫熒光
標(biāo)記方法：
AlexaFluor488 、CY2、 CY3、CY5、FITC
包埋方法：
樹脂包埋
成像平臺(tái)：
BioMapping 5000
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Fig. 4. Fluorescent images of Lectin-DyLight 594 labeled vasculature in the brain tissue after Lowicryl HM20 resin embedding. (a) Maximum intensity projections of a 20-μm-thick coronal slice. (b-d) Corresponding magnification of regions indicated in (a). (e-g) High magnification images of the boxed regions in (b-d), respectively. All images were acquired using 20 × (NA = 1.0), at a 0.42 × 0.42 × 1.00 μm 3 voxel size on a confocal microscope (LSM780, ZEISS). Scale bars: (a) 1 mm; (b-d) 100 μm; (e-g) 30 μm.
Fig. 5. Imaging large volume immunolabeled mouse brain tissue after Lowicryl HM20 resin embedding. (a) 3D presentation of the TH immunolabeled mouse brain block. Enlargements of the fine structures of TH-positive axonal fibers in the cortex (b) and TH-positive soma located in the thalamus (c). Images were acquired by successive high-resolution stage-scanning microscopy at a 0.16 × 0.16 × 1.00 μm 3 voxel size. (a) 6200 × 5000 × 1200 μm 3 ; (b) 480 × 480 × 380 μm 3 ; (c) 480 × 480 × 510 μm 3
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Fig. 4. Fluorescent images of Lectin-DyLight 594 labeled vasculature in the brain tissue after Lowicryl HM20 resin embedding. (a) Maximum intensity projections of a 20-μm-thick coronal slice. (b-d) Corresponding magnification of regions indicated in (a). (e-g) High magnification images of the boxed regions in (b-d), respectively. All images were acquired using 20 × (NA = 1.0), at a 0.42 × 0.42 × 1.00 μm 3 voxel size on a confocal microscope (LSM780, ZEISS). Scale bars: (a) 1 mm; (b-d) 100 μm; (e-g) 30 μm.
Fig. 5. Imaging large volume immunolabeled mouse brain tissue after Lowicryl HM20 resin embedding. (a) 3D presentation of the TH immunolabeled mouse brain block. Enlargements of the fine structures of TH-positive axonal fibers in the cortex (b) and TH-positive soma located in the thalamus (c). Images were acquired by successive high-resolution stage-scanning microscopy at a 0.16 × 0.16 × 1.00 μm 3 voxel size. (a) 6200 × 5000 × 1200 μm 3 ; (b) 480 × 480 × 380 μm 3 ; (c) 480 × 480 × 510 μm 3
2017年7月10日，華中科技大學(xué)武漢光電國(guó)家研究中心曾紹群教授課題組，提出了一種結(jié)合大樣本樹脂包埋和 iDISCO 免疫熒光染色的方法來獲得高空間分辨率的生物分子的方法。 這一方法結(jié)合fMOST技術(shù)，能夠以高分辨率成像免疫標(biāo)記的大體積組織。文章發(fā)表在《Biomed Opt Express》雜志上。
參考文獻(xiàn)
參考文獻(xiàn)[1]:Gang Y, Liu X, Wang X, Zhang Q, Zhou H, Chen R, Liu L, Jia Y, Yin F, Rao G, Chen J, Zeng S. Plastic embedding immunolabeled large-volume samples for three-dimensional high-resolution imaging. Biomed Opt Express. (2017);8(8):3583-3596.
武漢東湖新技術(shù)開發(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