Characterization of an industry-grade CMOS camera well suited for single molecule localization microscopy \u2013 high performance super-resolution at low cost

Robin Diekmann,Matthias Simonis,Thomas Huser,Mark Schüttpelz,Marcel Müller,Katharina Till

doi:10.1038/s41598-017-14762-6

Robin Diekmann, Matthias Simonis + Show 4 more

Open Access

https://doi.org/10.1038/s41598-017-14762-6

Copy DOI

Journal: Scientific Reports	Publication Date: Oct 31, 2017
Citations: 41	License type: open-access

Affiliation: Bielefeld University, University of Oxford

Abstract

Many commercial as well as custom-built fluorescence microscopes use scientific-grade cameras that represent a substantial share of the instrument’s cost. This holds particularly true for super-resolution localization microscopy where high demands are placed especially on the detector with respect to sensitivity, noise, and also image acquisition speed. Here, we present and carefully characterize an industry-grade CMOS camera as a cost-efficient alternative to commonly used scientific cameras. Direct experimental comparison of these two detector types shows widely similar performance for imaging by single molecule localization microscopy (SMLM). Furthermore, high image acquisition speeds are demonstrated for the CMOS detector by ultra-fast SMLM imaging.

Highlights

The advance of multiple super-resolution fluorescence microscopy techniques[1], such as single molecule localization microscopy (SMLM), structured illumination microscopy (SIM)[2,3], stimulated emission depletion (STED)[4,5], and super-resolution fluctuation imaging (SOFI)[6] has had a huge impact on the life sciences within the last couple of years
Such a large FOV exceeds the illuminated area in most implementations for dSTORM imaging, though it has recently been demonstrated how to adapt the illumination schemes in order to use the large detector areas of modern scientific-grade complementary metal-oxide-semiconductor (sCMOS) cameras more efficiently[27,28,29]
Though some differences in their performance can be expected, the utilized first generation camera already shows superior performance when compared to the industry-grade complementary metal-oxide-semiconductor (CMOS) camera with respect to noise and quantum efficiency in the deep red spectral range

Summary

Introduction

The advance of multiple super-resolution fluorescence microscopy techniques[1], such as single molecule localization microscopy (SMLM), structured illumination microscopy (SIM)[2,3], stimulated emission depletion (STED)[4,5], and super-resolution fluctuation imaging (SOFI)[6] has had a huge impact on the life sciences within the last couple of years. Except for very low signal levels, higher localization precisions can be achieved[15], which is one of the major performance marks in SMLM This is complemented by the development of advanced fluorophores[20], superior imaging buffer compositions[21], as well as sophisticated approaches to tailoring the on-state time of fluorescent labels[22]. While Holm et al.[23] have recently begun to demonstrate the use of a standard charge-coupled device (CCD) camera in a cost-efficient SMLM setup, the use of an industry-grade complementary metal-oxide-semiconductor (CMOS) camera for this purpose was just demonstrated by Ma et al.[24]. The utilization of the industry-grade CMOS camera in standard setups can contribute to a considerable cost reduction in dSTORM and other SMLM techniques without remarkably compromising the resulting images

Methods

Results

Conclusion