Multiplexed PSF Engineering for Three-Dimensional Multicolor Particle Tracking

被引：18

作者：

Opatovski, Nadav ^{[1
]}

Ezra, Yael Shalev ^{[2
]}

Weiss, Lucien E. ^{[2
]}

Ferdman, Boris ^{[1
]}

Orange-Kedem, Reut ^{[1
]}

Shechtman, Yoav ^{[1
,2
]}

机构：

[1] Technion Israel Inst Technol, Russel Berrie Nanotechnol Inst, IL-3200003 Haifa, Israel

[2] Technion Israel Inst Technol, Dept Biomed Engn, IL-3200003 Haifa, Israel

来源：

NANO LETTERS | 2021年 / 21卷 / 13期

基金：

欧盟地平线“2020”; 欧洲研究理事会;

关键词：

microscopy; PSF engineering; super-resolution; single-particle tracking; multicolor imaging; SINGLE-MOLECULE SPECTROSCOPY; LOCALIZATION MICROSCOPY; DYNAMICS; REVEALS; MOTION; COLOR;

D O I：

10.1021/acs.nanolett.1c02068

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Three-dimensional spatiotemporal tracking of microscopic particles in multiple colors is a challenging optical imaging task. Existing approaches require a trade-off between photon efficiency, field of view, mechanical complexity, spectral specificity, and speed. Here, we introduce multiplexed point-spread-function engineering that achieves photon-efficient, 3D multicolor particle tracking over a large field of view. This is accomplished by first chromatically splitting the emission path of a microscope to different channels, engineering the point-spread function of each, and then recombining them onto the same region of the camera. We demonstrate our technique for simultaneously tracking five types of emitters in vitro as well as colocalization of DNA loci in live yeast cells.

引用

页码：5888 / 5895

页数：8

共 47 条

[1]

Abrahamsson S, 2013, NAT METHODS, V10, P60, DOI [10.1038/NMETH.2277, 10.1038/nmeth.2277]

[2] Correlations of three-dimensional motion of chromosomal loci in yeast revealed by the double-helix point spread function microscope [J].