Department of Optics and Quantum Electronics, University of Szeged, 6720, Szeged, Dóm tér 9, Hungary.
BRC Institute of Genetics, Biological Research Centre HAS, Szeged, Hungary.
Sci Rep. 2019 Jan 28;9(1):798. doi: 10.1038/s41598-018-37341-9.
Super-resolution localization microscopy provides a powerful tool to study biochemical mechanisms at single molecule level. Although the lateral position of the fluorescent dye molecules can be determined routinely with high precision, measurement of other modalities such as 3D and multicolor without the degradation of the original super-resolved image is still in the focus. In this paper a dual-objective multimodal single molecule localization microscopy (SMLM) technique has been developed, optimized and tested. The proposed optical arrangement can be implemented onto a conventional inverted microscope without serious system modification. The performance of the method was tested using fluorescence beads, F-actin filaments and sarcomere structures. It was shown that the proposed imaging method does not degrade the image quality of the original SMLM 2D image but could provide information on the axial position or emission spectra of the dye molecules.
超分辨率定位显微镜为研究单分子水平的生化机制提供了强大的工具。虽然可以常规地以高精度确定荧光染料分子的横向位置，但在不降低原始超分辨图像质量的情况下，对其他模式（如 3D 和多色）的测量仍然是焦点。本文提出并优化了一种双目标多模态单分子定位显微镜（SMLM）技术并进行了测试。该方法可以在不进行严重系统修改的情况下，在传统的倒置显微镜上实现。该方法的性能使用荧光珠、F-肌动蛋白丝和肌节结构进行了测试。结果表明，所提出的成像方法不会降低原始 SMLM 2D 图像的图像质量，但可以提供染料分子的轴向位置或发射光谱的信息。
