New Imaging Technique Based on Photoswitchable Raman Probe Developed by Researchers

A diagram illustrating the RESORT imaging system. The sample is labeled with photoswitchable Raman probes and then exposed to two-color infrared laser pulses, ultraviolet light, and a donut-shaped beam of visible light. These techniques limit the area where Raman scattering occurs, resulting in highly precise and detailed images. Credit: 2023 Ozeki et al

Advancements in imaging techniques have revolutionized the examination of biological samples at the microscopic level. Researchers from the University of Tokyo have taken a step further by combining elements from two leading imaging methods to create the RESORT imaging technique. This groundbreaking concept enables the observation of living systems with unparalleled detail. The results of this study can be found in the journal Science Advances.

Throughout history, optical devices have allowed us to explore the microscopic world with increasing precision. The more we can see, the more we can understand. Thus, there is a constant drive to improve our tools for examining the world around us, both externally and internally.

Modern microscopic imaging techniques have surpassed the capabilities of traditional microscopes. Two major technologies in this field are super-resolution fluorescence imaging, which offers high spatial resolution, and vibrational imaging, which allows for versatile labeling of various cellular components but compromises spatial resolution.

Professor Yasuyuki Ozeki from the University of Tokyo’s Research Center for Advanced Science and Technology explained the motivation behind the development of RESORT: “We aimed to overcome the limitations of existing imaging techniques and create something superior. RESORT, which stands for reversible saturable optical Raman transitions, combines the advantages of super-resolution fluorescence and vibrational imaging without the drawbacks of either. This laser-based technique utilizes Raman scattering, a unique interaction between light and molecules, to identify and analyze samples under a microscope. We successfully demonstrated RESORT imaging of mitochondria in cells, validating the effectiveness of this technique.”

New imaging technique is no last resort
A comparison between RESORT and the previous imaging technique known as stimulated Raman scattering (SRS). RESORT provides sharper images, enabling researchers to identify and label cellular components, such as mitochondria, with greater accuracy. Credit: 2023 Ozeki et al

The RESORT imaging process involves several stages, which, despite sounding complex, are less intricate than the techniques currently in use. First, the specific components of the sample to be imaged are labeled with photoswitchable Raman probes, which can control Raman scattering through the use of different laser lights employed by RESORT.

Next, the sample is placed within an optical setup that illuminates and captures an image of the sample. To facilitate this, the sample is irradiated with two-color infrared laser pulses for Raman scattering detection, ultraviolet light, and a donut-shaped beam of visible light. Together, these techniques confine Raman scattering to a specific area, allowing for highly precise imaging with exceptional spatial resolution.

According to Ozeki: “The goal is not just to obtain higher-resolution images of microscopic samples, as electron microscopes already excel in this area. However, electron microscopes tend to damage or disrupt the samples they observe. With RESORT, we are aiming to expand the palette of Raman probes to include more colors. This will allow us to capture the dynamics of various components within living samples, providing unprecedented insights into complex biological processes, disease mechanisms, and potential therapeutic interventions.”

While the primary focus of the research team was to enhance microscopic imaging in medical research and related fields, the laser design advancements made in the development of RESORT could have applications in other laser-related sectors that require high power or precise control, such as materials science.

More information:
Jingwen Shou et al, Super-resolution vibrational imaging based on photoswitchable Raman probe, Science Advances (2023). DOI: 10.1126/sciadv.ade9118. www.science.org/doi/10.1126/sciadv.ade9118

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University of Tokyo


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Researchers create new imaging technique based on photoswitchable Raman probe (2023, June 16)
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