小柯机器人

美国约翰霍普金斯医学院Jie Xiao研究团队发现，成对距离分布校正 (DDC) 算法能消除单分子定位显微镜 (SMLM) 中由闪烁引起的假阳性结果。相关论文于2021年5月31日发表在《自然-方法学》杂志上。

研究人员提出了一种称为距离分布校正 (DDC) 的算法，无需任何额外校准即可消除由闪烁引起的重复定位。该方法通过使用比平均荧光寿命更长的时间分隔来定位荧光间的距离，从成像序列中自然捕获不同荧光团之间真实的成对距离分布。

使用真实的成对分布，研究人员可以定义和最大可能的获得一组不包含闪烁假阳性的定位信息。DDC 在获取真实空间组织和荧光发射器数量的最接近估计方面取得了巨大的进步，在广泛的应用中能够准确地重建和量化SMLM图像。

据悉，SMLM依赖于荧光团的闪烁成像，即荧光亮和暗状态之间的随机切换。闪烁会产生来源于同一荧光团的多个定位，其可以造成定量分析混乱和难以解析的结果。

附：英文原文

Title: A pairwise distance distribution correction (DDC) algorithm to eliminate blinking-caused artifacts in SMLM

Author: Christopher H. Bohrer, Xinxing Yang, Shreyasi Thakur, Xiaoli Weng, Brian Tenner, Ryan McQuillen, Brian Ross, Matthew Wooten, Xin Chen, Jin Zhang, Elijah Roberts, Melike Lakadamyali, Jie Xiao

Issue&Volume: 2021-05-31

Abstract: Single-molecule localization microscopy (SMLM) relies on the blinking behavior of a fluorophore, which is the stochastic switching between fluorescent and dark states. Blinking creates multiple localizations belonging to the same fluorophore, confounding quantitative analyses and interpretations. Here we present a method, termed distance distribution correction (DDC), to eliminate blinking-caused repeat localizations without any additional calibrations. The approach relies on obtaining the true pairwise distance distribution of different fluorophores naturally from the imaging sequence by using distances between localizations separated by a time much longer than the average fluorescence survival time. We show that, using the true pairwise distribution, we can define and maximize the likelihood, obtaining a set of localizations void of blinking artifacts. DDC results in drastic improvements in obtaining the closest estimate of the true spatial organization and number of fluorescent emitters in a wide range of applications, enabling accurate reconstruction and quantification of SMLM images. Distance distribution correction (DDC) eliminates repeat localizations caused by fluorophore blinking without the need for calibrations. Use of DDC yields accurate and quantifiable single-molecule localization microscopy data.

DOI: 10.1038/s41592-021-01154-y

Source: https://www.nature.com/articles/s41592-021-01154-y

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex