Laemmel Elisabeth, Genet Magalie, Le Goualher Georges, Perchant Aymeric, Le Gargasson Jean-François, Vicaut Eric
Laboratoire d'Etude de la Microcirculation, Université Paris 7, France.
J Vasc Res. 2004 Sep-Oct;41(5):400-11. doi: 10.1159/000081209. Epub 2004 Sep 30.
This study investigated the capability of fibered confocal fluorescence microscopy (FCFM) to provide in vivo microvascular observations. FCFM is specifically designed for in vivo in situ observation thanks to a probe composed of a fiber bundle and micro-optics having a diameter as small as 650 microm. In the first part of the study, we compared the main characteristics of FCFM with those of intravital fluorescence microscopy (IFM). A mouse cremaster preparation was used as a common basis to allow for imaging with both modalities. We discussed the feasibility of obtaining quantitative measurements usually provided by IFM in the context of FCFM: morphometry, capillary permeability, functional capillary density, vasoconstriction and dilation effects. In addition, the possibility to visualize fluorescent red blood cells or leukocytes was also evaluated. Phototoxicity issues and limitations of FCFM were also discussed. We showed that FCFM allows observations and measurements usually provided by IFM and that the real-time capability of the system, as well as the flexibility and small diameter of the optical probe enable micro-invasiveness and can extend imaging capabilities for in vivo in situ observations when compared to IFM.
本研究调查了纤维共聚焦荧光显微镜(FCFM)进行体内微血管观察的能力。FCFM专门设计用于体内原位观察,这得益于其由一束光纤和直径小至650微米的微光学器件组成的探头。在研究的第一部分,我们将FCFM的主要特性与活体荧光显微镜(IFM)的特性进行了比较。使用小鼠提睾肌标本作为共同基础,以便两种方式都能进行成像。我们讨论了在FCFM的背景下获得通常由IFM提供的定量测量的可行性:形态测量、毛细血管通透性、功能性毛细血管密度、血管收缩和扩张效应。此外,还评估了可视化荧光红细胞或白细胞的可能性。还讨论了光毒性问题和FCFM的局限性。我们表明,FCFM能够进行通常由IFM提供的观察和测量,并且该系统的实时能力以及光学探头的灵活性和小直径使其具有微侵入性,与IFM相比,可以扩展体内原位观察的成像能力。