Scientists have introduced a novel microscopy method named BonFIRE that merges fluorescence microscopy with vibrational microscopy, allowing for the observation of biological processes at the individual molecule level. This pioneering technique enhances selectivity and sensitivity, granting researchers the ability to create images of molecules using vibrational contrast. Acknowledgment: Caltech
Caltech’s research team has crafted BonFIRE, a state-of-the-art microscopy approach that unifies fluorescence with vibrational microscopy. This innovative method delivers unmatched single-molecule imaging and employs isotopes to generate various vibrational hues, granting profound insight into biological molecules and mechanisms.
While most people envision observing glass slides with simple organisms through a microscope, microscopes have the capacity to reveal much more. A freshly devised type of microscopy from the California Institute of Technology (Caltech) facilitates the viewing of the actual molecules constituting living organisms.
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The BonFIRE Technique
As detailed in Nature Photonics, researchers from Lu Wei’s lab, an assistant professor of chemistry and researcher with the Heritage Medical Research Institute, unveil bond-selective fluorescence-detected infrared-excited spectro-microscopy, also known as BonFIRE.
BonFIRE integrates two microscopy methodologies, enhancing selectivity and sensitivity, and affording scientists the capacity to observe biological processes at the groundbreaking single-molecule level, thus comprehending biological functionalities at the molecular level.
According to study co-author Dongkwan Lee, a chemical engineering graduate student, “Our advanced microscope now permits us to view individual molecules with vibrational contrast, something existing technologies struggle to achieve.”
BonFIRE’s functioning is being demonstrated by postdoctoral scholar Haomin Wang (left) and graduate student Dongkwan Lee (right). Acknowledgment: Caltech
Techniques Behind BonFIRE
BonFIRE consists of fluorescence microscopy, which visualizes molecules by using fluorescent tags that glow when imaged, and vibrational microscopy that leverages natural bond vibrations within a molecule to identify it. The process, involving infrared light bombardment, allows the differentiation of different bonds in a manner analogous to recognizing different guitar strings by their tones.
Combining Strengths
Assistant professor Lu Wei notes that while fluorescence microscopy facilitates single-molecule observation, it lacks detailed chemical information, unlike vibrational microscopy, which provides such information but requires larger quantities of the molecule.
BonFIRE transcends these limitations by fusing vibrations with fluorescence, synergizing the advantages of both techniques. The mechanism involves staining the sample with a specific dye, followed by sequential bombardment with infrared and higher-energy light, enabling the microscope to capture images of whole cells or individual molecules.
Future Prospects
Study co-author Haomin Wang, a postdoctoral scholar research associate in chemistry, expresses the team’s fascination and enthusiasm for turning this spectroscopy process into a modern bioimaging tool. Over three years of development has led to their custom BonFIRE microscope, with optimization to achieve the current performance level.
The scientists in their publication also demonstrate the capability to mark biomolecules with “colors” by utilizing various isotopes of the dye’s constituent atoms. These isotopes result in different vibrational frequencies.
Wei elaborates that, unlike traditional fluorescence microscopy, BonFIRE employs infrared light to excite diverse chemical bonds, yielding a spectrum of vibrational colors. “You can tag and image multiple targets from the same sample simultaneously, exposing life’s molecular diversity in breathtaking detail. We anticipate showcasing the imaging capacity with numerous colors in live cells shortly.”
Reference: “Bond Selective Fluorescence Imaging with Single Molecule Sensitivity” by Haomin Wang, Dongkwan Lee, Yulu Cao, Xiaotian Bi, Jiajun Du, Kun Miao, and Lu Wei, 29 June 2023, Nature Photonics. DOI: 10.1038/s41566-023-01243-8
The research also includes additional chemistry graduate students Yulu Cao, Xiaotian Bi, Jiajun Du, and Kun Miao.
The National Institutes of Health and the Alfred P. Sloan Foundation provided financial support for the study.
Frequently Asked Questions (FAQs) about BonFIRE
What is the BonFIRE technique in microscopy?
BonFIRE is a cutting-edge microscopy technique developed by Caltech researchers. It combines fluorescence microscopy and vibrational microscopy, allowing scientists to visualize biological processes at the single-molecule level. This innovative method offers unparalleled sensitivity and selectivity and can be used to provide deep insights into biological molecules and processes.
How does BonFIRE microscopy differ from conventional methods?
Unlike traditional fluorescence microscopy, BonFIRE synergizes both fluorescence and vibrational microscopy, permitting researchers to view individual molecules with vibrational contrast. This enables richer chemical information and the ability to image whole cells or individual molecules, revealing life’s molecular diversity in stunning detail.
What are the main applications of the BonFIRE technique?
The main applications of the BonFIRE technique include single-molecule imaging, using isotopes to create various vibrational colors, and revealing deep insights into biological molecules and processes. It’s a promising tool for modern bioimaging that could significantly advance the study of molecular biology.
Who are the key researchers behind the development of BonFIRE?
The key researchers behind BonFIRE include the lab of Lu Wei, assistant professor of chemistry at Caltech, and associated scholars such as Haomin Wang and Dongkwan Lee, along with other graduate students.
How does the BonFIRE technique combine the strengths of fluorescence and vibrational microscopy?
BonFIRE combines the strengths of fluorescence and vibrational microscopy by coupling vibrations to fluorescence. It works by staining the sample with a fluorescent dye, followed by a bombardment of infrared light tuned to excite a specific bond found in the dye. A second higher-energy pulse of light causes it to fluoresce, allowing the microscope to image entire cells or single molecules.
What is the future prospect of the BonFIRE microscopy technique?
The future prospects of BonFIRE include the development of a novel tool for modern bioimaging and the ability to tag biomolecules with “colors” using various isotopes. Researchers are excited about turning the spectroscopy process into a bioimaging tool and hope to demonstrate the imaging capability with tens of colors in live cells in the near future.
More about BonFIRE
- Nature Photonics
- Caltech
- Heritage Medical Research Institute
- National Institutes of Health
- Alfred P. Sloan Foundation
