Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion

Yang Zhang, Ki Hee Song, Biqin Dong, Janel L. Davis, Guangbin Shao, Cheng Sun, Hao F Zhang*

*Corresponding author for this work

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We developed transmission diffraction grating-based spectroscopic single-molecule localization microscopy (sSMLM) to collect the spatial and spectral information of single-molecule blinking events concurrently. We characterized the spectral heterogeneities of multiple far-red emitting dyes in a high-throughput manner using sSMLM. We also investigated the influence of spectral dispersion on the single-molecule identification performance of fluorophores with large spectral overlapping. The careful tuning of spectral dispersion in grating-based sSMLM permitted simultaneous three-color super-resolution imaging in fixed cells with a single objective lens at a relatively low photon budget. Our sSMLM has a compact optical design and can be integrated with conventional localization microscopy to provide add-on spectroscopic analysis capability.

Original languageEnglish (US)
Pages (from-to)2248-2255
Number of pages8
JournalApplied optics
Volume58
Issue number9
DOIs
StatePublished - Jan 1 2019

Fingerprint

Microscopic examination
microscopy
Imaging techniques
Molecules
molecules
blinking
Spectroscopic analysis
Optical design
Fluorophores
Diffraction gratings
spectroscopic analysis
gratings (spectra)
budgets
Lenses
Photons
Dyes
Tuning
dyes
tuning
lenses

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Engineering (miscellaneous)
  • Electrical and Electronic Engineering

Cite this

Zhang, Yang ; Song, Ki Hee ; Dong, Biqin ; Davis, Janel L. ; Shao, Guangbin ; Sun, Cheng ; Zhang, Hao F. / Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion. In: Applied optics. 2019 ; Vol. 58, No. 9. pp. 2248-2255.
@article{94a2ab055b54450c8d1c2eec4a4444cd,
title = "Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion",
abstract = "We developed transmission diffraction grating-based spectroscopic single-molecule localization microscopy (sSMLM) to collect the spatial and spectral information of single-molecule blinking events concurrently. We characterized the spectral heterogeneities of multiple far-red emitting dyes in a high-throughput manner using sSMLM. We also investigated the influence of spectral dispersion on the single-molecule identification performance of fluorophores with large spectral overlapping. The careful tuning of spectral dispersion in grating-based sSMLM permitted simultaneous three-color super-resolution imaging in fixed cells with a single objective lens at a relatively low photon budget. Our sSMLM has a compact optical design and can be integrated with conventional localization microscopy to provide add-on spectroscopic analysis capability.",
author = "Yang Zhang and Song, {Ki Hee} and Biqin Dong and Davis, {Janel L.} and Guangbin Shao and Cheng Sun and Zhang, {Hao F}",
year = "2019",
month = "1",
day = "1",
doi = "10.1364/AO.58.002248",
language = "English (US)",
volume = "58",
pages = "2248--2255",
journal = "Applied Optics",
issn = "0003-6935",
publisher = "The Optical Society",
number = "9",

}

Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion. / Zhang, Yang; Song, Ki Hee; Dong, Biqin; Davis, Janel L.; Shao, Guangbin; Sun, Cheng; Zhang, Hao F.

In: Applied optics, Vol. 58, No. 9, 01.01.2019, p. 2248-2255.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion

AU - Zhang, Yang

AU - Song, Ki Hee

AU - Dong, Biqin

AU - Davis, Janel L.

AU - Shao, Guangbin

AU - Sun, Cheng

AU - Zhang, Hao F

PY - 2019/1/1

Y1 - 2019/1/1

N2 - We developed transmission diffraction grating-based spectroscopic single-molecule localization microscopy (sSMLM) to collect the spatial and spectral information of single-molecule blinking events concurrently. We characterized the spectral heterogeneities of multiple far-red emitting dyes in a high-throughput manner using sSMLM. We also investigated the influence of spectral dispersion on the single-molecule identification performance of fluorophores with large spectral overlapping. The careful tuning of spectral dispersion in grating-based sSMLM permitted simultaneous three-color super-resolution imaging in fixed cells with a single objective lens at a relatively low photon budget. Our sSMLM has a compact optical design and can be integrated with conventional localization microscopy to provide add-on spectroscopic analysis capability.

AB - We developed transmission diffraction grating-based spectroscopic single-molecule localization microscopy (sSMLM) to collect the spatial and spectral information of single-molecule blinking events concurrently. We characterized the spectral heterogeneities of multiple far-red emitting dyes in a high-throughput manner using sSMLM. We also investigated the influence of spectral dispersion on the single-molecule identification performance of fluorophores with large spectral overlapping. The careful tuning of spectral dispersion in grating-based sSMLM permitted simultaneous three-color super-resolution imaging in fixed cells with a single objective lens at a relatively low photon budget. Our sSMLM has a compact optical design and can be integrated with conventional localization microscopy to provide add-on spectroscopic analysis capability.

UR - http://www.scopus.com/inward/record.url?scp=85063112142&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85063112142&partnerID=8YFLogxK

U2 - 10.1364/AO.58.002248

DO - 10.1364/AO.58.002248

M3 - Article

VL - 58

SP - 2248

EP - 2255

JO - Applied Optics

JF - Applied Optics

SN - 0003-6935

IS - 9

ER -