Ultrafast Widefield Mid-Infrared Photothermal Heterodyne Imaging.

Ultrafast Widefield Mid-Infrared Photothermal Heterodyne Imaging.
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DOI:
10.1021/acs.analchem.2c02548
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发表时间:
2022-10-18
影响因子:
7.4
通讯作者:
Schmidt, Florian M.
Schmidt, Florian M.
中科院分区:
化学1区
文献类型:
--
作者:
Paiva, Eduardo M.;Schmidt, Florian M.

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中红外光热显微镜(MIP)是一种具有中红外衍射极限外高空间分辨率的灵敏、快速化学成像的宝贵工具。采用光电探测器点扫描和锁定检测的外差MIP通常具有最高的灵敏度,而最快的系统使用基于相机的宽视场MIP和脉冲探针光。其中一个挑战是在大视场内同时实现高灵敏度、空间分辨率和速度。在这里,我们提出了宽视场中红外光热外差(WIPH)成像,其中数字频域锁定(DFdLi)滤波器用于同时多谐波解调由单个相机像素记录的MIP信号,帧率高达200 kHz。DFdLi滤波器支持使用连续波探测光,从而消除了对同步方案的需求,并允许测量MIP衰减曲线。WIPH方法的特点是成像铁氰化钾微粒,并应用于检测3T3-L1成纤维细胞中的脂滴(炔-棕榈酸),在2100 cm-1左右的细胞沉默光谱区域使用外腔量子级联激光器。该系统在128 × 128 μm视场中以5.52的信噪比和1 μm的空间分辨率实现了每秒4000幅WIPH图像。该技术为生物、医学和材料科学中快速过程的实时化学成像开辟了道路。
Mid-infrared photothermal (MIP) microscopy is a valuable tool for sensitive and fast chemical imaging with high spatial resolution beyond the mid-infrared diffraction limit. The highest sensitivity is usually achieved with heterodyne MIP employing photodetector point-scans and lock-in detection, while the fastest systems use camera-based widefield MIP with pulsed probe light. One challenge is to simultaneously achieve high sensitivity, spatial resolution, and speed in a large field of view. Here, we present widefield mid-infrared photothermal heterodyne (WIPH) imaging, where a digital frequency-domain lock-in (DFdLi) filter is used for simultaneous multiharmonic demodulation of MIP signals recorded by individual camera pixels at frame rates up to 200 kHz. The DFdLi filter enables the use of continuous-wave probe light, which, in turn, eliminates the need for synchronization schemes and allows measuring MIP decay curves. The WIPH approach is characterized by imaging potassium ferricyanide microparticles and applied to detect lipid droplets (alkyne-palmitic acid) in 3T3-L1 fibroblast cells, both in the cell-silent spectral region around 2100 cm–1 using an external-cavity quantum cascade laser. The system achieved up to 4000 WIPH images per second at a signal-to-noise ratio of 5.52 and 1 μm spatial resolution in a 128 × 128 μm field of view. The technique opens up for real-time chemical imaging of fast processes in biology, medicine, and material science.
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