印度墨水灌注法重建小鼠全腦連續(xù)血管網(wǎng)
染色方法：
印度墨水染色
標(biāo)記方法：
印度墨水心臟灌注
包埋方法：
樹脂包埋
成像平臺：
BioMapping 5000
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Figure 1. Vessel acquisition and reconstruction in the whole mouse brain. A: The dorsal view of the gelatin-Indian ink perfused brain; after embedding with Spurr resin and imaging using MOST to acquire the three-dimensional cerebrovascular dataset, five 100- m m thick coronal planes were selected and MIP-reconstructed in the olfactory bulb (B), frontal (C), hippocampus (D), midbrain (E) and cerebellum (F); the results showed desirable perfusion in the whole brain, and no blank areas were found in the brain entity. G: ROI in Figure 1D; the capillary network was continuous, and no gaps were found. H: The left view of the whole brain; several major vessels were marked. I: The top view of the whole brain. cc: corpus callosum; hf: hippocampus; th: thalamus. B–F: Bar=1 mm; G: Bar=200 m m, H: Bar=1 mm.
Figure 2. Assessment of capillary continuity. Four ROIs (Columns A–D) with size of 20062006100 m m 3 at cortex, amygdaloid nucleus, thalamus, cerebellar lobule were selected and used to analyze the vascular continuity. The four ROIs were indicated sequentially with white arrows in Figure 1D, F and G. ROI A was taken as an example. On the top row, the MIP reconstruction of this ROI covered with vascular tracking results by using Amira was shown. The vascular terminal could be located through vascular tracking results projection on xy, yz and xz orthogonal planes (Rows 2–4). The vascular terminals inside the ROI were marked with red balls, while the terminals on the boundary were marked with blue balls. The panels on the bottom showed the details around the vascular terminals, most of which were caused by caliber narrowing.
Figure 3. Blood supply pathway tracking and analysis in the right thalamus. A: The right view of the brain. A clear blood pathway begins in the VA, passes through the BA and the thalamoperforating artery, enters the thalamus, and finally drains out through the thalamostriate vein, the great cerebral vein of Galen and the transverse sinus. B: The vessels with diameters larger than 5 m m in the thalamus, red for arteries and blue for veins. C: A vascular segment from the thalamoperforating artery to the thalamostriate vein through the capillary, colored by diameter. D: The capillaries in the pathway from the artery to the vein. E–F: ROIs from D and C, showing the narrowing in the capillaries. * Identifies the vascular narrowing sites in C, E and F.
Video S1 Blood supply pathway tracking in the thalamus.
Video S2 Slices appear in a sequential way to show 100 um MIP reconstructions on coronal plane for the whole brain.
Video S3 Slices appear in a sequential way to show 100 um MIP reconstructions on sagittal plane for the whole brain.
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Figure 1. Vessel acquisition and reconstruction in the whole mouse brain. A: The dorsal view of the gelatin-Indian ink perfused brain; after embedding with Spurr resin and imaging using MOST to acquire the three-dimensional cerebrovascular dataset, five 100- m m thick coronal planes were selected and MIP-reconstructed in the olfactory bulb (B), frontal (C), hippocampus (D), midbrain (E) and cerebellum (F); the results showed desirable perfusion in the whole brain, and no blank areas were found in the brain entity. G: ROI in Figure 1D; the capillary network was continuous, and no gaps were found. H: The left view of the whole brain; several major vessels were marked. I: The top view of the whole brain. cc: corpus callosum; hf: hippocampus; th: thalamus. B–F: Bar=1 mm; G: Bar=200 m m, H: Bar=1 mm.
Figure 2. Assessment of capillary continuity. Four ROIs (Columns A–D) with size of 20062006100 m m 3 at cortex, amygdaloid nucleus, thalamus, cerebellar lobule were selected and used to analyze the vascular continuity. The four ROIs were indicated sequentially with white arrows in Figure 1D, F and G. ROI A was taken as an example. On the top row, the MIP reconstruction of this ROI covered with vascular tracking results by using Amira was shown. The vascular terminal could be located through vascular tracking results projection on xy, yz and xz orthogonal planes (Rows 2–4). The vascular terminals inside the ROI were marked with red balls, while the terminals on the boundary were marked with blue balls. The panels on the bottom showed the details around the vascular terminals, most of which were caused by caliber narrowing.
Figure 3. Blood supply pathway tracking and analysis in the right thalamus. A: The right view of the brain. A clear blood pathway begins in the VA, passes through the BA and the thalamoperforating artery, enters the thalamus, and finally drains out through the thalamostriate vein, the great cerebral vein of Galen and the transverse sinus. B: The vessels with diameters larger than 5 m m in the thalamus, red for arteries and blue for veins. C: A vascular segment from the thalamoperforating artery to the thalamostriate vein through the capillary, colored by diameter. D: The capillaries in the pathway from the artery to the vein. E–F: ROIs from D and C, showing the narrowing in the capillaries. * Identifies the vascular narrowing sites in C, E and F.
Video S1 Blood supply pathway tracking in the thalamus.
Video S2 Slices appear in a sequential way to show 100 um MIP reconstructions on coronal plane for the whole brain.
Video S3 Slices appear in a sequential way to show 100 um MIP reconstructions on sagittal plane for the whole brain.
2014年1月30日，華中科技大學(xué)武漢光電國家研究中心李安安教授實驗組，使用MOST技術(shù)，結(jié)合改進的明膠-印度墨汁血管灌注，展示了小鼠全腦血管網(wǎng)絡(luò)成像。本研究為同時研究小鼠腦的整個宏觀和微觀血管網(wǎng)絡(luò)提供了一種有效的方法。文章發(fā)表在《PLoS One》雜志上。
參考文獻
參考文獻[1]:Xue S, Gong H, Jiang T, Luo W, Meng Y, Liu Q, Chen S, Li A. Indian-ink perfusion based method for reconstructing continuous vascular networks in whole mouse brain. PLoS One (2014);9(1):e88067.
武漢東湖新技術(shù)開發(fā)區(qū)豹澥街道高科園三路9號武漢光谷精準(zhǔn)醫(yī)療產(chǎn)業(yè)基地2.1期13棟12層