Laboratory for Fluorescence Dynamics, University of California - Biomedical Engineering, 3210 Natural Sciences 2, Irvine, California 92672, USA.
J Biophotonics. 2011 Jun;4(6):415-24. doi: 10.1002/jbio.201100002. Epub 2011 Apr 1.
We developed an optical imaging method based on a feedback principle in which the specific scan pattern is adapted according to the shape of the sample. The feedback approach produces nanometer-resolved 3D images of very small and moving features in live cells in seconds. We show images of microvilli in live cultured opossum kidney cells expressing NaPi co-transporter proteins with different GFP constructs and images of cell protrusions in a collagen matrix with a resolution of about 20 nm. We found that in the microvilli the NaPi proteins can be found clustered. Along cell protrusions in 3D we identified cellular adhesions to the extracellular matrix. Our approach to super-resolution and to 3D nanoimaging is different than other proposed methods that break the diffraction limit using non-linear effects or are based on single molecule localization.
我们开发了一种基于反馈原理的光学成像方法，该方法根据样品的形状来适应特定的扫描模式。反馈方法可以在几秒钟内生成活细胞中非常小且移动的特征的纳米分辨率 3D 图像。我们展示了用不同 GFP 构建体表达 NaPi 共转运蛋白的活培养袋状肾细胞中的微绒毛的图像，以及胶原基质中分辨率约为 20nm 的细胞突起的图像。我们发现，在微绒毛中，NaPi 蛋白可以聚集在一起。沿着 3D 的细胞突起，我们鉴定了细胞与细胞外基质的黏附。我们的超分辨率和 3D 纳米成像方法与其他提出的方法不同，其他方法使用非线性效应突破衍射极限，或者基于单分子定位。
