近日，法国巴黎文理研究大学Ludovic Jullien等研究人员合作发现，荧光可测量光强度。这一研究成果于2023年11月23日在线发表在《自然—方法学》期刊上。

研究人员表示，尽管许多科学学科都需要对光照强度进行定量测量，但现有的荧光显微镜样品光剂量测量技术却无法同时检索光照强度以及各种波长和强度的空间分布。

为了解决这一局限性，研究人员开发了两种快速、直接的方案，其使用有机染料和荧光蛋白作为感光计。第一种方案依赖于其荧光强度在恒定光照下以单指数方式衰减和/或上升的分子系统。第二种方案依靠一种具有宽吸收性的光化学惰性荧光团来反向计算从一个波长到另一个波长的光强度。为了证明其用途，这些方案被用于定量描述各种荧光成像系统的光空间分布，以及校准市售仪器和光源的照明。

附：英文原文

Title: Fluorescence to measure light intensity

Author: Lahlou, Alinor, Tehrani, Hessam Sepasi, Coghill, Ian, Shpinov, Yuriy, Mandal, Mrinal, Plamont, Marie-Aude, Aujard, Isabelle, Niu, Yuxi, Nedbal, Ladislav, Lazr, Dusan, Mahou, Pierre, Supatto, Willy, Beaurepaire, Emmanuel, Eisenmann, Isabelle, Desprat, Nicolas, Croquette, Vincent, Jeanneret, Raphal, Le Saux, Thomas, Jullien, Ludovic

Issue&Volume: 2023-11-23

Abstract: Despite the need for quantitative measurements of light intensity across many scientific disciplines, existing technologies for measuring light dose at the sample of a fluorescence microscope cannot simultaneously retrieve light intensity along with spatial distribution over a wide range of wavelengths and intensities. To address this limitation, we developed two rapid and straightforward protocols that use organic dyes and fluorescent proteins as actinometers. The first protocol relies on molecular systems whose fluorescence intensity decays and/or rises in a monoexponential fashion when constant light is applied. The second protocol relies on a broad-absorbing photochemically inert fluorophore to back-calculate the light intensity from one wavelength to another. As a demonstration of their use, the protocols are applied to quantitatively characterize the spatial distribution of light of various fluorescence imaging systems, and to calibrate illumination of commercially available instruments and light sources.

DOI: 10.1038/s41592-023-02063-y

Source: https://www.nature.com/articles/s41592-023-02063-y