Deep UV autofluorescence microscopy for cell biology and tissue histology

Abstract

In this work we demonstrate that DUV autofluorescence is a powerful tool for tissue histology and cell biology.

Deep UV excited muscle cell autofluorescence varies with the fibre type

“…17,25,37,38 In the literature, fluorescence emission around 410 nm is usually assigned to collagen or elastin, 25 but these proteins are specific to the extracellular space, and are completely absent in the intracellular space where the measurements were made. Irrespective of the fibre type and cell localization, all spectra show the same shape (see The spectrum shows two peaks at 332 and 410 nm with a shoulder at 302 nm.…”

“…Insight into the muscle fibre composition is of great importance in a wide range of scientific fields, such as clinical research, biomechanics, and muscle food science. 24,25 Like front-face fluorescence, DUV microspectroscopy requires no external specific probes or labelling, instead it allows the use of the intrinsic fluorescence that many biomolecules display when excited at wavelengths below 350 nm. 13,14 However, the use of monoclonal antibodies (mAbs) against myosin heavy chain (MyHC) iso-forms enabled a more precise classification of fibres I, IIA, IIX and IIB.…”

“…24,25 The DISCO beamline of the SOLEIL synchrotron has developed a DUV fluorescence microscope coupled to a synchrotron beamline, providing fine-tuneable excitation from 180 to 600 nm and full-spectrum acquisition from each point scanned, to study DUV-excited fluorescence emitted from nanovolumes directly inside live cells or tissue biopsies. 24,25 The DISCO beamline of the SOLEIL synchrotron has developed a DUV fluorescence microscope coupled to a synchrotron beamline, providing fine-tuneable excitation from 180 to 600 nm and full-spectrum acquisition from each point scanned, to study DUV-excited fluorescence emitted from nanovolumes directly inside live cells or tissue biopsies.…”

Molecular changes in gelatin aging observed by NIR and fluorescence spectroscopy

“…The strong fluorescence intensity at 305 nm under a 280 nm excitation is assigned to tyrosine (Chagnot et al, 2015;Jamme et al, 2013;Lakowicz, 2006;Theron et al, 2014). The decrease in fluorescence intensity of tyrosine during aging can be linked to a change in its molecular environment (Lakowicz, 2006) or to a decrease in tyrosine content owing to its degradation or to chemical reaction such as dityrosine bond formation.…”

Deep-UV biological imaging by lanthanide ion molecular protection

“…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. Although 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. Suppressing the molecular photodegradation is essential for unlocking these limitations in the practical use of DUV light for biological imaging.…”