Villiger M, Goulley J, Friedrich M, Grapin-Botton A, Meda P, Lasser T, Leitgeb R A
Laboratoire d'Optique Biomédicale, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Diabetologia. 2009 Aug;52(8):1599-607. doi: 10.1007/s00125-009-1383-y. Epub 2009 May 30.
AIMS/HYPOTHESIS: Structural and functional imaging of the islets of Langerhans and the insulin-secreting beta cells represents a significant challenge and a long-lasting objective in diabetes research. In vivo microscopy offers a valuable insight into beta cell function but has severe limitations regarding sample labelling, imaging speed and depth, and was primarily performed on isolated islets lacking native innervations and vascularisation. This article introduces extended-focus optical coherence microscopy (xfOCM) to image murine pancreatic islets in their natural environment in situ, i.e. in vivo and in a label-free condition.
Ex vivo measurements on excised pancreases were performed and validated by standard immunohistochemistry to investigate the structures that can be observed with xfOCM. The influence of streptozotocin on the signature of the islets was investigated in a second step. Finally, xfOCM was applied to make measurements of the murine pancreas in situ and in vivo.
xfOCM circumvents the fundamental physical limit that trades lateral resolution for depth of field, and achieves fast volumetric imaging with high resolution in all three dimensions. It allows label-free visualisation of pancreatic lobules, ducts, blood vessels and individual islets of Langerhans ex vivo and in vivo, and detects streptozotocin-induced islet destruction.
CONCLUSIONS/INTERPRETATION: Our results demonstrate the potential value of xfOCM in high-resolution in vivo studies to assess islet structure and function in animal models of diabetes, aiming towards its use in longitudinal studies of diabetes progression and islet transplants.
目的/假设:对胰岛和分泌胰岛素的β细胞进行结构和功能成像,是糖尿病研究中的一项重大挑战和长期目标。体内显微镜检查为β细胞功能提供了有价值的见解,但在样品标记、成像速度和深度方面存在严重限制,且主要是在缺乏天然神经支配和血管化的分离胰岛上进行。本文介绍了扩展聚焦光学相干显微镜(xfOCM),用于在自然环境中对小鼠胰腺胰岛进行原位成像,即在体内且无标记条件下成像。
对切除的胰腺进行离体测量,并通过标准免疫组织化学进行验证,以研究xfOCM可观察到的结构。第二步研究链脲佐菌素对胰岛特征的影响。最后,应用xfOCM对小鼠胰腺进行原位和体内测量。
xfOCM规避了以横向分辨率换取景深的基本物理限制,并在所有三个维度上实现了具有高分辨率的快速体积成像。它能够在离体和体内对胰腺小叶、导管、血管和单个胰岛进行无标记可视化,并检测链脲佐菌素诱导的胰岛破坏。
结论/解读:我们的结果证明了xfOCM在高分辨率体内研究中评估糖尿病动物模型中胰岛结构和功能的潜在价值,旨在将其用于糖尿病进展和胰岛移植的纵向研究。