Photon-free (s)CMOS camera characterization for artifact reduction in high- and super-resolution microscopy

Diekmann，Robin1,3; Deschamps，Joran1,4; Li，Yiming1,5; Deguchi，Takahiro1; Tschanz，Aline1,2; Kahnwald，Maurice1,6; Matti，Ulf1,7; Ries，Jonas1

2022-12-01

10.1038/s41467-022-30907-2

Nature Communications   影响因子和分区

2041-1723

Modern implementations of widefield fluorescence microscopy often rely on sCMOS cameras, but this camera architecture inherently features pixel-to-pixel variations. Such variations lead to image artifacts and render quantitative image interpretation difficult. Although a variety of algorithmic corrections exists, they require a thorough characterization of the camera, which typically is not easy to access or perform. Here, we developed a fully automated pipeline for camera characterization based solely on thermally generated signal, and implemented it in the popular open-source software Micro-Manager and ImageJ/Fiji. Besides supplying the conventional camera maps of noise, offset and gain, our pipeline also gives access to dark current and thermal noise as functions of the exposure time. This allowed us to avoid structural bias in single-molecule localization microscopy (SMLM), which without correction is substantial even for scientific-grade, cooled cameras. In addition, our approach enables high-quality 3D super-resolution as well as live-cell time-lapse microscopy with cheap, industry-grade cameras. As our approach for camera characterization does not require any user interventions or additional hardware implementations, numerous correction algorithms that rely on camera characterization become easily applicable.

SCI

英语

NI论文

其他

FP7 Ideas: European Research Council[ERC CoG-724489];Human Frontier Science Program[RGY0065/2017];Foundation for the National Institutes of Health[U01 EB021223];

Science & Technology - Other Topics

Multidisciplinary Sciences

WOS:000809574500008

NATURE PORTFOLIO

2-s2.0-85131805039

Scopus

1.Cell Biology and Biophysics Unit,European Molecular Biology Laboratory (EMBL),Heidelberg,Germany
2.Collaboration for Joint PhD Degree Between EMBL and Heidelberg University,Faculty of Biosciences,Heidelberg,Germany
3.LaVision Biotec GmbH,Bielefeld,Germany
4.Fondazione Human Technopole,Milan,Italy
5.Department of Biomedical Engineering,Southern University of Science and Technology,Shenzhen,China
6.Friedrich Miescher Institute for Biomedical Research,Basel,Switzerland
7.Abberior Instruments GmbH,Göttingen,Germany

Diekmann，Robin,Deschamps，Joran,Li，Yiming,et al. Photon-free (s)CMOS camera characterization for artifact reduction in high- and super-resolution microscopy[J]. Nature Communications,2022,13(1).

Diekmann，Robin.,Deschamps，Joran.,Li，Yiming.,Deguchi，Takahiro.,Tschanz，Aline.,...&Ries，Jonas.(2022).Photon-free (s)CMOS camera characterization for artifact reduction in high- and super-resolution microscopy.Nature Communications,13(1).

Diekmann，Robin,et al."Photon-free (s)CMOS camera characterization for artifact reduction in high- and super-resolution microscopy".Nature Communications 13.1(2022).