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用于细胞成像的基于超导转变边缘传感器的少光子光谱共聚焦显微镜

Few-Photon Spectral Confocal Microscopy for Cell Imaging Using Superconducting Transition Edge Sensor.

作者信息

Niwa Kazuki, Hattori Kaori, Fukuda Daiji

机构信息

Research Institute for Physical Measurement, National Metrology Institute of Japan, National Institute of Advanced Industrial Science and Technology (AIST), Tokyo, Japan.

AIST-UTokyo Advanced Operando-Measurement Technology Open Innovation Laboratory, Kashiwa, Japan.

出版信息

Front Bioeng Biotechnol. 2021 Dec 15;9:789709. doi: 10.3389/fbioe.2021.789709. eCollection 2021.

Abstract

A superconducting transition edge sensor (TES) is an energy-dispersive single-photon detector that distinguishes the wavelength of each incident photon from visible to near-infrared (NIR) without using spectral dispersive elements. Here, we introduce an application of the TES technique for confocal laser scanning microscopy (CLSM) as proof of our concept of ultra-sensitive and wide-band wavelength range color imaging for biological samples. As a reference sample for wide-band observation, a fixed fluorescence-labeled cell sample stained with three different color dyes was observed using our TES-based CLSM method. The three different dyes were simultaneously excited by irradiating 405 and 488 nm lasers, which were coupled using an optical fiber combiner. Even when irradiated at low powers of 80 and 120 nW with the 405 and 488 nm lasers respectively, emission signals were spectrally detected by the TES and categorized into four wavelength bands: up to 500 nm (blue), from 500 to 600 nm (green), from 600 to 800 nm (red), and from 800 to 1,200 nm (NIR). Using a single scan, an RGB color image and an NIR image of the fluorescent cell sample were successfully captured with tens of photon signals in a 40 ms exposure time for each pixel. This result demonstrates that TES is a useful wide-band spectral photon detector in the field of life sciences.

摘要

超导转变边缘传感器(TES)是一种能量色散单光子探测器,它无需使用光谱色散元件就能区分从可见光到近红外(NIR)的每个入射光子的波长。在此,我们介绍了TES技术在共聚焦激光扫描显微镜(CLSM)中的应用,以证明我们对生物样品进行超灵敏和宽带波长范围彩色成像的概念。作为宽带观察的参考样品,使用我们基于TES的CLSM方法观察了用三种不同颜色染料染色的固定荧光标记细胞样品。通过照射405和488nm激光同时激发这三种不同的染料,这两种激光使用光纤合束器进行耦合。即使分别以80和120nW的低功率照射405和488nm激光,TES仍能对发射信号进行光谱检测,并将其分为四个波长带:500nm以下(蓝色)、500至600nm(绿色)、600至800nm(红色)以及800至1200nm(近红外)。通过单次扫描,在每个像素40ms的曝光时间内成功捕获了荧光细胞样品的RGB彩色图像和近红外图像,其中包含数十个光子信号。这一结果表明,TES是生命科学领域中一种有用的宽带光谱光子探测器。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a294/8715037/eb9e572086e1/fbioe-09-789709-g001.jpg

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