Abstract Interference Reflection Microscopy (IRM) is an optical technique that relies on the interference between the reflected light from an incident beam as it passes through materials of different refractive indices. This technique has been successfully used to image microtubules, biologically important biofilaments with a diameter of 25 nm. However, it is often desirable to image both the microtubule and microtubule interacting proteins simultaneously. Here we present a simple modification to a standard multicolour total internal reflection fluorescence (TIRF) microscope that enables simultaneous high‐speed IRM and single molecule TIRF imaging. Our design utilises a camera for each channel (IRM and TIRF) allowing independent optimisation of camera parameters for the two different modalities. We illustrate its application by imaging unlabelled microtubules and GFP‐labelled end‐binding protein EB1, which forms comets on the tips of polymerising microtubules. Our design is easily implemented, and with minimal cost, making it accessible to any laboratory with an existing fluorescence microscope.

Jeffrey O. Spector

Jiayi Chen

Ewa Szczesna

Antonina Roll‐Mecak

Journal of Microscopy

《显微镜杂志》是致力于显微镜科学的最古老的期刊，也是皇家显微镜学会唯一经同行评审的出版物。它发表的论文报告了显微镜技术的最新发展，例如显微镜技术的进步或新的应用领域。该杂志不寻求发表显微镜或标本制备的常规应用，即使提交的材料可能具有很高的科学价值。范围涵盖物理和生物科学的研究，并涵盖使用光、电子、X 射线和其他方法的成像方法辐射以及原子力和近场技术。欢迎跨学科研究。与显微镜有关的论文也很受欢迎，涉及光学理论、光谱学、新的标本制备和操作方法以及图像记录、处理和分析，包括活体标本的动态分析。出版类型包括全文、热门话题快速通信和评论文章。考虑提交评论文章的作者应首先联系编辑部。

The Journal of Microscopy is the oldest journal dedicated to the science of microscopy and is the only peer-reviewed publication of the Royal Microscopical Society. It publishes papers reporting the latest developments in microscopy techniques, including advancements in microscopy technology and new areas of application. While the journal does not seek to publish routine applications of microscopy or specimen preparation, submissions with high scientific value will be considered. The scope encompasses research in both physical and biological sciences, covering imaging methods using light, electron, X-ray, and other radiation techniques, as well as atomic force and near-field techniques. Interdisciplinary research is also welcomed. Papers related to microscopy, particularly those involving optical theory, spectroscopy, new specimen preparation and operational methods, as well as image recording, processing, and analysis (including dynamic analysis of live specimens), are highly encouraged. Publication types include full articles, rapid communications on hot topics, and review articles. Authors considering submitting review articles should first contact the editorial office.

《Journal of Microscopy》是显微镜科学领域最古老的期刊，也是皇家显微镜学会唯一经过同行评审的出版物。该期刊发表关于显微镜技术最新发展的论文，包括显微镜技术的进步和新的应用领域。虽然该杂志不寻求发表显微镜或标本制备的常规应用，但即使提交的材料具有很高的科学价值，也会受到审查。期刊的范围涵盖物理和生物科学的研究，涉及使用光学、电子、X 射线及其他成像方法的辐射，以及原子力和近场技术。跨学科研究也受到欢迎。与显微镜相关的论文，尤其是涉及光学理论、光谱学、新的标本制备和操作方法，以及图像记录、处理和分析（包括活体标本的动态分析）也很受欢迎。出版类型包括全文、热门话题快速通信和评论文章。考虑提交评论文章的作者应首先联系编辑部。

Category	Quartile
工程技术	4
工程技术, 显微镜技术	4

Multicamera simultaneous total internal reflection and interference reflection microscopy

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