Abstract
Deep-UV (DUV) light is a sensitive probe for biological molecules such as nucleobases and aromatic amino acids due to specific absorption. However, the use of DUV light for imaging is limited because DUV can destroy or denature target molecules in a sample. Here we show that trivalent ions in the lanthanide group can suppress molecular photodegradation under DUV exposure, enabling a high signal-to-noise ratio and repetitive DUV imaging of nucleobases in cells. Underlying mechanisms of the photodegradation suppression can be excitation relaxation of the DUV-absorptive molecules due to energy transfer to the lanthanide ions, and/or avoiding ionization and reactions with surrounding molecules, including generation of reactive oxygen species, which can modify molecules that are otherwise transparent to DUV light. This approach, directly removing excited energy at the fundamental origin of cellular photodegradation, indicates an important first step towards the practical use of DUV imaging in a variety of biological applications.
Highlights
Deep-UV (DUV) light, corresponding to a wavelength range of 200-300 nm, is a sensitive probe of biological molecules which absorb DUV light remarkably well, but do not interact significantly with visible light, such as nucleobases, aromatic amino acids, and dopamine
We show that trivalent ions in the lanthanide group can suppress molecular photodegradation under DUV exposure, enabling a high signal-to-noise ratio and repetitive DUV imaging of nucleobases in cells
A variety of advanced DUV imaging techniques measuring absorption [22,23,24], resonance Raman scattering [25], fluorescence [26,27,28,29,30], and photoacoustic signals [31,32] of DUV-absorptive molecules have been developed recently, the destructive nature of DUV light limits some unique benefits of these techniques in exploiting biological molecules, which could be measured with high sensitivity if the photodegradation could be avoided
Summary
Deep-UV (DUV) light, corresponding to a wavelength range of 200-300 nm, is a sensitive probe of biological molecules which absorb DUV light remarkably well, but do not interact significantly with visible light, such as nucleobases, aromatic amino acids, and dopamine. DUV light has been used for label-free biomolecular detection in chromatography [1] and medical dialysis [2]. It has been employed for precise analysis of local structures of macromolecules, such as proteins [3,4,5,6,7,8] and nucleic acids [9], by DUV spectroscopy. DUV light can destroy or denature biological molecules due to absorption [10,11,12,13,14,15,16,17,18,19,20,21], which restricts its use in quantitative, repetitive, and/or high signal-to-noise ratio (SNR) analyses of target molecules in a limited detection volume by DUV imaging, especially for samples involving trace amounts. A variety of advanced DUV imaging techniques measuring absorption [22,23,24], resonance Raman scattering [25], fluorescence [26,27,28,29,30], and photoacoustic signals [31,32] of DUV-absorptive molecules have been developed recently, the destructive nature of DUV light limits some unique benefits of these techniques in exploiting biological molecules, which could be measured with high sensitivity if the photodegradation could be avoided
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