トラック&トレース法で顕微鏡の最大解像度を予測する(Track-and-trace method predicts best possible resolution in microscopy)

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2022-06-21 オランダ・デルフト工科大学(TUDelft)

デルフト工科大学の科学者は、超解像顕微鏡の限界についての洞察を提供し、最大解像度を決定するための新しい計算方法を提供します。研究チームによって以前行われた大きな精度向上は、彼らの発表によってニュアンスが変わりました。この技術は、生きている細胞内のプロセスを研究し、病気の原因を発見し、新しい薬を開発するために重要です。さらに、彼らの発表は、他の研究者が以前主張した大きな精度向上のニュアンスを含んでいます。今回の研究成果は、『Biophysical Journal』に掲載されました。

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反復変調強化単一分子局在顕微鏡法における精度 Precision in iterative modulation enhanced single-molecule localization microscopy

DylanKalisvaart,JelmerCnossen,Shih-TeHung,SjoerdStallinga,MichelVerhaegen,Carlas S.Smith
Biophysical Journal  Available online:25 May 2022
DOI:https://doi.org/10.1016/j.bpj.2022.05.027

Abstract

Modulation enhanced single-molecule localization microscopy (meSMLM) methods improve the localization precision by using patterned illumination to encode additional position information. Iterative meSMLM (imeSMLM) methods iteratively generate prior information on emitter positions, used to locally improve the localization precision during subsequent iterations. The Cramér-Rao lower bound cannot incorporate prior information to bound the best achievable localization precision because it requires estimators to be unbiased. By treating estimands as random variables with a known prior distribution, the Van Trees inequality (VTI) can be used to bound the best possible localization precision of imeSMLM methods. An imeSMLM method is considered, where the positions of in-plane standing-wave illumination patterns are controlled over the course of multiple iterations. Using the VTI, we analytically approximate a lower bound on the maximum localization precision of imeSMLM methods that make use of standing-wave illumination patterns. In addition, we evaluate the maximally achievable localization precision for different illumination pattern placement strategies using Monte Carlo simulations. We show that in the absence of background and under perfect modulation, the information content of signal photons increases exponentially as a function of the iteration count. However, the information increase is no longer exponential as a function of the iteration count under non-zero background, imperfect modulation, or limited mechanical resolution of the illumination positioning system. As a result, imeSMLM with two iterations reaches at most a fivefold improvement over SMLM at 8 expected background photons per pixel and 95% modulation contrast. Moreover, the information increase from imeSMLM is balanced by a reduced signal photon rate. Therefore, SMLM outperforms imeSMLM when considering an equal measurement time and illumination power per iteration. Finally, the VTI is an excellent tool for the assessment of the performance of illumination control and is therefore the method of choice for optimal design and control of imeSMLM methods.

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