Three-dimensional optical method for integrated visualization of mouse islet microstructure and vascular network with subcellular-level resolution.
Microscopic visualization of islets of Langerhans under normal and diabetic conditions is essential for understanding the pathophysiology of the disease. The intrinsic opacity of pancreata, however, limits optical accessibility for high-resolution light microscopy of islets in situ. Because the standard microtome-based, 2-D tissue analysis confines visualization of the islet architecture at a specific cut plane, 3-D representation of image data is preferable for islet assessment. We applied optical clearing to minimize the random light scattering in the mouse pancreatic tissue. The optical-cleared pancreas allowed penetrative, 3-D microscopic imaging of the islet microstructure and vasculature. Specifically, the islet vasculature was revealed by vessel painting-lipophilic dye labeling of blood vessels-for confocal microscopy. The voxel-based confocal micrographs were digitally processed with projection algorithms for 3-D visualization. Unlike the microtome-based tissue imaging, this optical method for penetrative imaging of mouse islets yielded clear, continuous optical sections for an integrated visualization of the islet microstructure and vasculature with subcellular-level resolution. We thus provide a useful imaging approach to change our conventional planar view of the islet structure into a 3-D panorama for better understanding of the islet physiology.
Fu
Ya-Yuan
YY
National Tsing Hua University, Department of Chemical Engineering, Hsinchu, Taiwan.
Lu
Chih-Hsuan
CH
Lin
Chi-Wen
CW
Juang
Jyuhn-Huarng
JH
Enikolopov
Grigori
G
Sibley
Eric
E
Chiang
Ann-Shyn
AS
Tang
Shiue-Cheng
SC
D000465:Algorithms; D000818:Animals; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007515:Islets of Langerhans / Q000098:blood supply* / Q000166:cytology*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D055806:Microvessels / Q000648:ultrastructure*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013347:Subcellular Fractions / Q000648:ultrastructure*; D018511:Systems Integration
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