Advanced Photonics, Volume. 7, Issue 3, 036003(2025)

Breaking the diffraction limit in molecular imaging by structured illumination mid-infrared photothermal microscopy

Pengcheng Fu1、†, Bo Chen1, Yongqing Zhang1, Liangyi Chen2, Hyeon Jeong Lee3,4、*, and Delong Zhang1、*
Author Affiliations
  • 1Zhejiang University, School of Physics, Zhejiang Key Laboratory of Micro-nano Quantum Chips and Quantum Control, Hangzhou, China
  • 2Peking University, Institute of Molecular Medicine, School of Future Technology, Peking-Tsinghua Center for Life Sciences, New Cornerstone Science Laboratory, State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Beijing Laboratory of Biomedical Imaging, Beijing, China
  • 3Zhejiang University, College of Biomedical Engineering and Instrument Science, Key Laboratory for Biomedical Engineering of Ministry of Education, Hangzhou, China
  • 4Zhejiang University, MOE Frontier Science Center for Brain Science & Brain-Machine Integration, Hangzhou, China
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    Figures & Tables(4)
    Principle of SIMIP. (a) Schematic of the SIMIP system. (b) Schematic diagram of illumination pattern on the imaging plane. (c) Isotropic extension of OTF by varying the phase and the orientation of the illumination pattern. The red dot represents the projection of the sample in the reciprocal space. (d) Simulated PSF for widefield imaging and SIM imaging, respectively. (e) Modulation of the fluorophore emission intensity by the IR pulse vibrational absorption of nearby molecules. (f) Chemical information obtained using SIMIP.
    Setup and data acquisition process of SIMIP. (a) Setup of SIMIP. SMF, single-mode fiber; SLM, spatial light modulator; HWP, half-wave plate; PBS, polarizing beam splitter; L, lens; M, mirrors; OAP, off-axis parabolic mirror; OBJ, objective; DM, dichroic mirror; sCMOS, scientific complementary metal-oxide semiconductor; QCL, quantum cascade laser. (b) The time scheme of SIMIP. (c) The process of obtaining SIMIP spectra.
    SIMIP imaging performance. (a) SIMIP images of 200-nm PMMA beads at 1730 cm−1. (b) Spectra of PMMA film measured by FTIR (blue) and PMMA beads measured by SIMIP (red). int., intensity; abs. absorption; a.u., arbitrary units. (c), (d) SIMIP images of 200-nm PMMA beads at different wavenumbers. (e), (f) The enlarged images of the selected region in panel (a) using SIMIP (e) and F-MIP/F-PTIR (f), respectively. (g) The intensity profiles of a single bead using SIMIP (red) and F-MIP/F-PTIR (black), respectively. (h) The intensity profiles of the PMMA beads images indicated by the white arrows in panels (e) and (f). (i), (j) The frequency domain amplitude images of the same beads using SIMIP (i) and F-MIP/F-PTIR (j). The yellow boxes in panels (i) and (j) highlight the same region.
    Fluorescence and SIMIP images of the mixture of PS beads and PMMA beads. (a), (b) Fluorescence (a) and SIMIP (b) images from the same position in the sample. (c), (d) The enlarged images of the selected region in panels (a) and (b). (e), (f) Standard FTIR spectra of PS (e) and PMMA samples (f), respectively. Vertical lines indicate the selected wavelengths.
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    Pengcheng Fu, Bo Chen, Yongqing Zhang, Liangyi Chen, Hyeon Jeong Lee, Delong Zhang, "Breaking the diffraction limit in molecular imaging by structured illumination mid-infrared photothermal microscopy," Adv. Photon. 7, 036003 (2025)

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    Paper Information

    Category: Research Articles

    Received: Dec. 2, 2024

    Accepted: Mar. 10, 2025

    Posted: Mar. 10, 2025

    Published Online: Apr. 14, 2025

    The Author Email: Jeong Lee Hyeon (hjlee@zju.edu.cn), Zhang Delong (dlzhang@zju.edu.cn)

    DOI:10.1117/1.AP.7.3.036003

    CSTR:32187.14.1.AP.7.3.036003

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