Abstract
Ultraviolet (UV) spectroscopy is a powerful tool for quantitative (bio)chemical analysis, but its application to molecular imaging and microscopy has been limited. Here we introduce ultraviolet hyperspectral interferometric (UHI) microscopy, which leverages coherent detection of optical fields to overcome significant challenges associated with UV spectroscopy when applied to molecular imaging. We demonstrate that this method enables quantitative spectral analysis of important endogenous biomolecules with subcellular spatial resolution and sensitivity to nanometer-scaled structures for label-free molecular imaging of live cells.
Highlights
The use of deep-UV light for microscopy offers many potential advantages over traditional methods, including higher spatial resolution due to the light’s shorter wavelength; and, when combined with spectroscopy, quantitative information with access to many endogenous molecules that play an important role in cell function and structure[1]
ultraviolet hyperspectral interferometric (UHI) microscopy leverages these recent advances along with coherent detection to overcome all of the aforementioned limitations of molecular imaging in the deep-UV region of the spectrum
With Fourier domain (FD) detection, this configuration yields an interferometric signal with high fringe visibility, and access to the complex optical fields across a large spectral region (e.g., 240 nm–450 nm)
Summary
The use of deep-UV light for microscopy offers many potential advantages over traditional methods, including higher spatial resolution due to the light’s shorter wavelength; and, when combined with spectroscopy, quantitative information with access to many endogenous molecules that play an important role in cell function and structure[1]. UHI microscopy leverages these recent advances along with coherent detection to overcome all of the aforementioned limitations of molecular imaging in the deep-UV region of the spectrum. With Fourier domain (FD) detection (i.e., light is detected as a function of wavelength), this configuration yields an interferometric signal with high fringe visibility, and access to the complex optical fields across a large spectral region (e.g., 240 nm–450 nm). This complex information allows us to correct for the chromatic aberration in the system by digitally refocusing. UHI microscopy enables deep-UV, wideband, high-resolution spectroscopic measurements, with high spatial resolution and sensitivity to nanometer-scaled spatial fluctuations
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