FRET-LPTEM for In-situ Imaging of Chemical Systems

doi:10.1007/s40242-025-4241-7

高等学校化学研究 ›› 2025, Vol. 41 ›› Issue (2): 319-325.doi: 10.1007/s40242-025-4241-7

FRET-LPTEM for In-situ Imaging of Chemical Systems

XU Zhun, ZHANG Deyi, XIONG Tianyu, WANG Huan

Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Key Laboratory of Polymer Chemistry and Physics, National Biomedical Imaging Center, Peking University, Beijing 100871, P. R. China

收稿日期:2024-12-14 接受日期:2025-01-25 出版日期:2025-04-01 发布日期:2025-03-31
通讯作者: WANG Huan,wanghuan_ccme@pku.edu.cn E-mail:wanghuan_ccme@pku.edu.cn
基金资助:
This work was supported by the Natural Science Foundation of Beijing Municipality, China (No. Z240010) and the National Natural Science Foundation of China (Nos. 22174006, T2495223). We thank Prof. SHAO Yuanhua for supporting the CO2-laser-based puller (P-2000, Sutter Instrument Co.) in making nanopipette liquid cells.

FRET-LPTEM for In-situ Imaging of Chemical Systems

XU Zhun, ZHANG Deyi, XIONG Tianyu, WANG Huan

Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Key Laboratory of Polymer Chemistry and Physics, National Biomedical Imaging Center, Peking University, Beijing 100871, P. R. China

Received:2024-12-14 Accepted:2025-01-25 Online:2025-04-01 Published:2025-03-31
Contact: WANG Huan,wanghuan_ccme@pku.edu.cn E-mail:wanghuan_ccme@pku.edu.cn
Supported by:
This work was supported by the Natural Science Foundation of Beijing Municipality, China (No. Z240010) and the National Natural Science Foundation of China (Nos. 22174006, T2495223). We thank Prof. SHAO Yuanhua for supporting the CO2-laser-based puller (P-2000, Sutter Instrument Co.) in making nanopipette liquid cells.

摘要/Abstract

摘要： An in-situ double-tilt holder has been made to integrate laser illumination and fluorescence-based spectroscopic analysis for conducting liquid-phase electron microscopy (LP-TEM) experiments using ordinary TEM. The setup differs from the existing geometry. Laser illumination and the collection of fluorescence signals were achieved using a single optical fiber, with efficiency optimized by adjusting the fiber position and grid tilt angle. Fluorescence emission of common organic dyes, propidium iodide (PI) and cyanine dyes, and Förster resonance energy transfer (FRET) signals of a FRET pair, Cy3/Cy5, were obtained from three types of liquid cells, including carbon film, graphene, and nanopipette liquid cells. The successful application of FRET-LPTEM enables LP-TEM experiments to be equipped with controlled light-triggering capability, detection of fluorogenic small molecules during chemical reactions, and the standard FRET experiments for macromolecules being conducted with LP-TEM. FRET-LPTEM presents opportunities for unraveling pathways underpinning the synthesis and assembly of optically active organic and biological materials.

关键词: In-situ double-tilt holder, Optical fiber, Light-coupling experiment, Förster resonance energy transfer (FRET), Liquidphase electron microscopy (LP-TEM)

Abstract: An in-situ double-tilt holder has been made to integrate laser illumination and fluorescence-based spectroscopic analysis for conducting liquid-phase electron microscopy (LP-TEM) experiments using ordinary TEM. The setup differs from the existing geometry. Laser illumination and the collection of fluorescence signals were achieved using a single optical fiber, with efficiency optimized by adjusting the fiber position and grid tilt angle. Fluorescence emission of common organic dyes, propidium iodide (PI) and cyanine dyes, and Förster resonance energy transfer (FRET) signals of a FRET pair, Cy3/Cy5, were obtained from three types of liquid cells, including carbon film, graphene, and nanopipette liquid cells. The successful application of FRET-LPTEM enables LP-TEM experiments to be equipped with controlled light-triggering capability, detection of fluorogenic small molecules during chemical reactions, and the standard FRET experiments for macromolecules being conducted with LP-TEM. FRET-LPTEM presents opportunities for unraveling pathways underpinning the synthesis and assembly of optically active organic and biological materials.

Key words: In-situ double-tilt holder, Optical fiber, Light-coupling experiment, Förster resonance energy transfer (FRET), Liquidphase electron microscopy (LP-TEM)

XU Zhun, ZHANG Deyi, XIONG Tianyu, WANG Huan. FRET-LPTEM for In-situ Imaging of Chemical Systems[J]. 高等学校化学研究, 2025, 41(2): 319-325.

XU Zhun, ZHANG Deyi, XIONG Tianyu, WANG Huan. FRET-LPTEM for In-situ Imaging of Chemical Systems[J]. Chemical Research in Chinese Universities, 2025, 41(2): 319-325.

参考文献

[1] Klinger M., J. Appl. Crystallogr., 2017, 50, 1226.
[2] Zou X., Hovmoller S., Acta Crystallogr. A, 2008, 64, 149.
[3] Li R., Li Z., Dong Z., Khor K. A., Crystals, 2016, 6, 105.
[4] Harris J. R., Arch. Biochem. Biophys., 2015, 581, 3.
[5] Shen P. S., Anal. Bioanal. Chem., 2018, 410, 2053.
[6] Hu H., Yang R., Zeng Z., ACS Nano, 2024, 18, 12598.
[7] Orji N. G., Badaroglu M., Barnes B. M., Beitia C., Bunday B. D., Celano U., Kline R. J., Neisser M., Obeng Y., Vladar A. E., Nat. Electron., 2018, 1, 532.
[8] Zhu Y., Zhao H., He Y., Wang R., J. Phys. D: Appl. Phys., 2021, 54, 443002.
[9] Han Y., Wang L., Cao K., Zhou J., Zhu Y., Hou Y., Lu Y., Chem. Rev., 2023, 123, 14119.
[10] Li K. Q., Bu Y. Q., Wang H. T., Front. Mater., 2023, 10, 1207024.
[11] Li J.-Y., Zhang D.-Y., Mao S., Wang H., Chin. J. Chem., 2023, 41, 679.
[12] Li J.-Y., Sun H., Wang H., Trends Chem., 2024, 6, 281.
[13] Li J.-Y., Liu F., Xu J., Kim Y.-J., Kwon O.-H., Xia B., Wang H., Granick S., Proc. Natl. Acad. Sci. USA, 2024, 121, e2314797121.
[14] Zhang D., Shao Y., Zhou J., Zhan Q., Wen Z., Mao S., Wei J., Qi L., Shao Y., Wang H., Proc. Natl. Acad. Sci. USA, 2024, 121, e2314320121.
[15] Wang H., Xu Z., Mao S., Granick S., ACS Nano, 2022, 16, 18526.
[16] McKinney S. A., Joo C., Ha T., Biophys. J., 2006, 91, 1941.
[17] Balci H., Arslan S., Myong S., Lohman T. M., Ha T., Biophys. J., 2011, 101, 976.
[18] Dong H., Xu F., Sun Z., Wu X., Zhang Q., Zhai Y., Tan X. D., He L., Xu T., Zhang Z., Duan X., Sun L., Nat. Nanotechnol., 2019, 14, 950.
[19] Gao P., Wang Z. Z., Liu K. H., Xu Z., Wang W. L., Bai X. D., Wang E. G., J. Mater. Chem., 2009, 19, 1002.
[20] Ohno Y., Taishi T., Yonenaga I., Phys. Status Solidi A, 2009, 206, 1904.
[21] Ohno Y., Appl. Phys. Express, 2012, 5, 125204.
[22] Cavalca F., Laursen A. B., Kardynal B. E., Dunin-Borkowski R. E., Dahl S., Wagner J. B., Hansen T. W., Nanotechnology, 2012, 23, 075705.
[23] Peng S., Wang Y., Braun M., Yin Y., Meng A. C., Tan W., Saini B., Severson K., Marshall A. F., Sytwu K., Baniecki J. D., Dionne J., Cai W., McIntyre P. C., Matter, 2023, 6, 2052.
[24] Duan T., Wang W., Cai S., Zhou Y., ACS Energy Lett., 2023, 8, 3048.
[25] Ohno Y., Yonenaga I., Appl. Surf. Sci., 2014, 302, 29.
[26] Yoshida K., Yamasaki J., Tanaka N., Appl. Phys. Lett., 2004, 84, 2542.
[27] Lu Y., Yin W.-J., Peng K.-L., Wang K., Hu Q., Selloni A., Chen F.-R., Liu L.-M., Sui M.-L., Nat. Commun., 2018, 9, 2752.
[28] Yu S., Jiang Y., Sun Y., Gao F., Zou W., Liao H., Dong L., Appl. Catal. B-Environ., 2021, 284, 119743.
[29] Weng B., Jiang Y., Liao H.-G., Roeffaers M. B. J., Lai F., Huang H., Tang Z., Nano Res., 2021, 14, 2805.
[30] Shindo D., Takahashi K., Murakami Y., Yamazaki K., Deguchi S., Suga H., Kondo Y., J. Electron Microsc. (Tokyo), 2009, 58, 245.
[31] Dombi P., Pápa Z., Vogelsang J., Yalunin S. V., Sivis M., Herink G., Schäfer S., Groß P., Ropers C., Lienau C., Rev. Mod. Phys., 2020, 92, 025003.
[32] Fu X., Liu S., Chen B., Tang J., Zhu Y., ACS Nano, 2021, 15, 6801.
[33] Zonnevylle A. C., Van Tol R. F. C., Liv N., Narvaez A. C., Effting A. P. J., Kruit P., Hoogenboom J. P., J. Microsc., 2013, 252, 58.
[34] Maruyama Y., Ebihara T., Nishiyama H., Suga M., Sato C., J. Struct. Biol., 2012, 180, 259.
[35] Berger C., Dumoux M., Glen T., Yee N. B. Y., Mitchels J. M., Patáková Z., Darrow M. C., Naismith J. H., Grange M., Nat. Commun., 2023, 14, 629.
[36] Mohammadian S., Agronskaia A. V., Blab G. A., van Donselaar E. G., de Heus C., Liv N., Klumperman J., Gerritsen H. C., Ultramicroscopy, 2020, 215, 113007.
[37] Agronskaia A. V., Valentijn J. A., van Driel L. F., Schneijdenberg C. T. W. M., Humbel B. M., van Bergen en Henegouwen P. M. P., Verkleij A. J., Koster A. J., Gerritsen H. C., J. Struct. Biol., 2008, 164, 183.
[38] Liu C., Ma C., Xu J., Qiao R., Sun H., Li X., Xu Z., Gao P., Wang E., Liu K., Bai X., Rev. Sci. Instrum., 2021, 92, 013704.
[39] Żak A. M., Nano Lett., 2022, 22, 9219.
[40] Fernando J. F. S., Zhang C., Firestein K. L., Golberg D., Small, 2017, 13, 1701564.
[41] Yu S., Jiang Y., Sun Y., Gao F., Zou W., Liao H., Dong L., Applied Catalysis B: Environmental, 2021, 284, 119743.
[42] Iqbal A., Arslan S., Okumus B., Wilson T. J., Giraud G., Norman D. G., Ha T., Lilley D. M. J., Proc. Natl. Acad. Sci. USA, 2008, 105, 11176.
[43] Roy R., Hohng S., Ha T., Nature Methods, 2008, 5, 507.
[44] Ashoka A. H., Aparin I. O., Reisch A., Klymchenko A. S., Chem. Soc. Rev., 2023, 52, 4525.
[45] Christie J. M., Arvai A. S., Baxter K. J., Heilmann M., Pratt A. J., O'Hara A., Kelly S. M., Hothorn M., Smith B. O., Hitomi K., Jenkins G. I., Getzoff E. D., Science, 2012, 335, 1492.
[46] Chang R., Zhao L., Xing R., Li J., Yan X., Chem. Soc. Rev., 2023, 52, 2688.
[47] Singh R., Wang Z., Marques C., Min R., Zhang B., Kumar S., Biosens. Bioelectron., 2023, 236, 115424.
[48] Park S., Rim S., Kim J. W., Park J., Sohn I.-B., Lee B. H., Sensors, 2018, 18, 4150.
[49] Kubelka J., Photochem. Photobiol. Sci., 2009, 8, 499.

FRET-LPTEM for In-situ Imaging of Chemical Systems

FRET-LPTEM for In-situ Imaging of Chemical Systems

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 3

编辑推荐

Metrics

本文评价

[1]	LIU Chunyu, WANG Shaoyan, FU Cuicui, LI Haibo, XU Shuping, XU Weiqing. Preparation of Surface-enhanced Raman Scattering(SERS)-active Optical Fiber Sensor by Laser-induced Ag Deposition and Its Application in Bioidentification of Biotin/Avidin[J]. 高等学校化学研究, 2015, 31(1): 25-30.
[2]	LI Ying-xiu, ZHU Lian-de, ZHU Guo-yi. A Novel Flow Injection Optical Fiber Biosensor for Hypoxanthine Based on Luminol Electrochemiluminescence[J]. 高等学校化学研究, 2003, 19(2): 240-244.
[3]	WANG Ying, LIU Wan-hui  . A Fluorescence Optical Fiber Sensor for Amrinon Using a New Oxazole Compound[J]. 高等学校化学研究, 1999, 15(4): 322-328.