荧光标记光纤极化效应的表征与校正

IF 1.5 4区工程技术 Q3 MICROSCOPY

Journal of microscopy Pub Date : 2024-04-29 DOI:10.1111/jmi.13308

Nandini Aggarwal, Richard Marsh, Stefania Marcotti, Tanya J Shaw, Brian Stramer, Susan Cox, Siân Culley

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引用次数: 0

摘要

摘要许多生物结构都以纤维和细丝的形式存在，对纤维组织的定量分析对于了解它们在正常生理条件和疾病中的功能都非常重要。为了使这些结构可视化，可以对纤维进行荧光标记和成像，并采用专门的图像分析方法来量化纤维排列的程度和强度。在这里，我们展示了荧光标记的纤维可显示极化发射，这种效应的强度因结构和荧光团特性而异。这会使纤维排列的自动分析产生偏差，并掩盖真实的潜在结构组织。我们提出了一种无需偏振分辨显微镜即可量化和校正这些偏振效应的方法，并展示了该方法在应用于荧光标记胶原凝胶图像时的功效，使纤维微观结构的表征更加可靠。

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Characterisation and correction of polarisation effects in fluorescently labelled fibres

SummaryMany biological structures take the form of fibres and filaments, and quantitative analysis of fibre organisation is important for understanding their functions in both normal physiological conditions and disease. In order to visualise these structures, fibres can be fluorescently labelled and imaged, with specialised image analysis methods available for quantifying the degree and strength of fibre alignment. Here we show that fluorescently labelled fibres can display polarised emission, with the strength of this effect varying depending on structure and fluorophore identity. This can bias automated analysis of fibre alignment and mask the true underlying structural organisation. We present a method for quantifying and correcting these polarisation effects without requiring polarisation‐resolved microscopy and demonstrate its efficacy when applied to images of fluorescently labelled collagen gels, allowing for more reliable characterisation of fibre microarchitecture.

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来源期刊

Journal of microscopy 工程技术-显微镜技术

CiteScore

4.30

自引率

5.00%

发文量

审稿时长

1 months

期刊介绍： The Journal of Microscopy is the oldest journal dedicated to the science of microscopy and the only peer-reviewed publication of the Royal Microscopical Society. It publishes papers that report on the very latest developments in microscopy such as advances in microscopy techniques or novel areas of application. The Journal does not seek to publish routine applications of microscopy or specimen preparation even though the submission may otherwise have a high scientific merit. The scope covers research in the physical and biological sciences and covers imaging methods using light, electrons, X-rays and other radiations as well as atomic force and near field techniques. Interdisciplinary research is welcome. Papers pertaining to microscopy are also welcomed on optical theory, spectroscopy, novel specimen preparation and manipulation methods and image recording, processing and analysis including dynamic analysis of living specimens. Publication types include full papers, hot topic fast tracked communications and review articles. Authors considering submitting a review article should contact the editorial office first.