Berguiga L, Roland T, Monier K, Elezgaray J, Argoul F
USR3010, UMR 5672, CNRS, Ecole Normale Supérieure de Lyon, Lyon, France.
Opt Express. 2011 Mar 28;19(7):6571-86. doi: 10.1364/OE.19.006571.
Imaging cellular internal structure at nanometer scale axial resolution with non invasive microscopy techniques has been a major technical challenge since the nineties. We propose here a complement to fluorescence based microscopies with no need of staining the biological samples, based on a Scanning Surface Plasmon Microscope (SSPM). We describe the advantages of this microscope, namely the possibility of both amplitude and phase imaging and, due to evanescent field enhancement by the surface plasmon resonance, a very high resolution in Z scanning (Z being the axis normal to the sample). We show for fibroblast cells (IMR90) that SSPM offers an enhanced detection of index gradient regions, and we conclude it is very well suited to discriminate regions of variable density in biological media such as cell compartments, nucleus, nucleoli and membranes.
自九十年代以来,利用非侵入性显微镜技术在纳米尺度轴向分辨率下对细胞内部结构进行成像一直是一项重大技术挑战。我们在此提出一种基于扫描表面等离子体显微镜(SSPM)的荧光显微镜补充方法,无需对生物样品进行染色。我们描述了这种显微镜的优点,即具有幅度和相位成像的可能性,并且由于表面等离子体共振增强了倏逝场,在Z扫描(Z为垂直于样品的轴)中具有非常高的分辨率。我们对成纤维细胞(IMR90)的研究表明,SSPM能够增强对折射率梯度区域的检测,并且我们得出结论,它非常适合区分生物介质中密度可变的区域,如细胞区室、细胞核、核仁和膜。