Nanotheranostics 2022; 6(1):50-61. doi:10.7150/ntno.63222 This issue


Review on Optical Imaging Techniques for Multispectral Analysis of Nanomaterials

Haeni Lee1,*, Jaeheung Kim1,*, Hyung-Hoi Kim2,✉, Chang-Seok Kim1,✉, Jeesu Kim1,✉

1. Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University, Busan 46241, Republic of Korea.
2. Department of Laboratory Medicine and Biomedical Research Institute, Pusan National University Hospital and Pusan National University School of Medicine, Busan 49241, Republic of Korea.
*These authors contributed equally to this work.

This is an open access article distributed under the terms of the Creative Commons Attribution License ( See for full terms and conditions.
Lee H, Kim J, Kim HH, Kim CS, Kim J. Review on Optical Imaging Techniques for Multispectral Analysis of Nanomaterials. Nanotheranostics 2022; 6(1):50-61. doi:10.7150/ntno.63222. Available from

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Graphic abstract

Biomedical imaging is an essential tool for investigating biological responses in vivo. Among the several imaging techniques, optical imaging systems with multispectral analysis of nanoparticles have been widely investigated due to their ability to distinguish the substances in biological tissues in vivo. This review article focus on multispectral optical imaging techniques that can provide molecular functional information. We summarize the basic principle of the spectral unmixing technique that enables the delineation of optical chromophores. Then, we explore the principle, typical system configuration, and biomedical applications of the representative optical imaging techniques, which are fluorescence imaging, two-photon microscopy, and photoacoustic imaging. The results in the recent studies show the great potential of the multispectral analysis techniques for monitoring responses of biological systems in vivo.

Keywords: Multispectral imaging, Nanoparticles, Spectral unmixing, Contrast-enhanced imaging, Biomedical imaging