Video-rate Nanoscopy Using SCMOS Camera-specific Single-molecule Localization Algorithms

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2013 May 28

PMID 23708387

Citations 215

Authors

Fang Huang

Tobias M P Hartwich

Felix E Rivera-Molina

Yu Lin

Whitney C Duim

Jane J Long

Pradeep D Uchil

Jordan R Myers

Michelle A Baird

Walther Mothes

Michael W Davidson

Derek Toomre

Joerg Bewersdorf

Affiliations

Soon will be listed here.

Abstract

Newly developed scientific complementary metal-oxide semiconductor (sCMOS) cameras have the potential to dramatically accelerate data acquisition, enlarge the field of view and increase the effective quantum efficiency in single-molecule switching nanoscopy. However, sCMOS-intrinsic pixel-dependent readout noise substantially lowers the localization precision and introduces localization artifacts. We present algorithms that overcome these limitations and that provide unbiased, precise localization of single molecules at the theoretical limit. Using these in combination with a multi-emitter fitting algorithm, we demonstrate single-molecule localization super-resolution imaging at rates of up to 32 reconstructed images per second in fixed and living cells.

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