High-resolution three-dimensional imaging of islet-infiltrate interactions based on optical projection tomography assessments of the intact adult mouse pancreas.
A predicament when assessing the mechanisms underlying the pathogenesis of type-1 diabetes (T1D) has been to maintain simultaneous global and regional information on the loss of insulin-cell mass and the progression of insulitis. We present a procedure for high-resolution 3-D analyses of regions of interest (ROIs), defined on the basis of global assessments of the 3-D distribution, size, and shape of molecularly labeled structures within the full volume of the intact mouse pancreas. We apply a refined protocol for optical projection tomography (OPT)-aided whole pancreas imaging in combination with confocal laser scanning microscopy of site-directed pancreatic microbiopsies. As such, the methodology provides a useful tool for detailed cellular and molecular assessments of the autoimmune insulitis in T1D. It is anticipated that the same approach could be applied to other areas of research where 3-D molecular distributions of both global and regional character is required.
Alanentalo
Tomas
T
Umeå University, Umeå Centre for Molecular Medicine, S-901 87, Umeå, Sweden.
Lorén
Christina E
CE
Larefalk
Asa
A
Sharpe
James
J
Holmberg
Dan
D
Ahlgren
Ulf
U
D000465:Algorithms; D000818:Animals; D005260:Female; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007515:Islets of Langerhans / Q000166:cytology*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D010179:Pancreas; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D041622:Tomography, Optical / Q000379:methods*