Fast epi-detected broadband multiplex CARS and SHG imaging of mouse skull cells

Coherent Raman microscopy has become a powerful tool in label-free, non-destructive and fast cell imaging. Here we apply high spectral resolution multiplex coherent anti-Stokes Raman scattering (MCARS) microspectroscopy in the high wavenumber region to the study of the cell cycle. We show that heterochromatin - the condensed state of chromatin - can be visualised by means of the vibrational signature of proteins taking part in its condensation. Thus, we are able to identify chromosomes and their movement during mitosis, as well as structures like nucleoli and nuclear border in interphase. Furthermore, the specific organization of the endoplasmic reticulum during mitosis is highlighted. Finally, we stress that MCARS can reveal the biochemical impact of the fixative method at the cellular level. Beyond the study of the cell cycle, this work introduces a label-free imaging approach that enables the visualization of cellular processes where chromatin undergoes rearrangements.

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“…The MCARS system is an adaptation, in forward configuration, of our epi-detected custom-built setup 42 . It is designed as follows.…”

Section: Methodsmentioning

confidence: 99%

Multiplex coherent anti-Stokes Raman scattering highlights state of chromatin condensation in CH region

Ben

Rajaofara

Couderc

et al. 2019

Sci Rep

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“…As such, design considerations to optimize narrow band spectral resolution and in vivo imaging speed were a priority. A number of broadband spectral NRI techniques have since been described (79)(80)(81)(82)(83), with a handful demonstrated in vivo (83)(84)(85)(86)(87)(88). The flexibility of the existing instrumentation on MANTIS readily allows us to integrate such broadband approaches into MANTIS with expanded optical configurations.…”

Section: Discussionmentioning

confidence: 99%

Multi-modal Nonlinear Optical and Thermal Imaging Platform for Label-Free Characterization of Biological Tissue

Adams

Mehl

Lieser

et al. 2020

Preprint

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14The ability to characterize the combined structural, functional, and thermal properties of 15 biophysically dynamic samples is needed to address critical questions related to tissue structure, 16physiological dynamics, and disease progression. Towards this, we have developed an imaging 17 platform that enables multiple nonlinear imaging modalities to be combined with thermal 18imaging on a common sample. Here we demonstrate label-free multimodal imaging of live cells, 19excised tissues, and live rodent brain models. While potential applications of this technology are 20 wide-ranging, we expect it to be especially useful in addressing biomedical research questions 21 aimed at the biomolecular and biophysical properties of tissue and their physiology. 22

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“…Conventional CARS systems use high numerical aperture (NA) objective lenses to converge the pump light and the Stokes light onto the samples. Such systems achieve the research on highly arti cialisolating individual cells and the quantitative characterization of intracellular biomolecules [6][7][8][9] . Although studying single cells is helpful, cells in biological tissues also need to be studied much because it is well known that cells behave quite differently in biological tissues than they do when separated in arti cial environments 10 .…”

Section: Introductionmentioning

confidence: 99%

Flexible Minimally Invasive CARS Measurement Method with Tapered Optical Fiber Probe for Single-Cell Application

Wang

Jiang

Liu

et al. 2022

Preprint

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We proposed and demonstrated a flexible, endoscopic, and minimally invasive coherent anti-Raman Stokes scattering (CARS) measurement method for single-cell application, employing a tapered optical fiber probe. A few-mode fiber (FMF), whose generated four-wave mixing band is out of CARS signals, was selected to fabricate tapered optical fiber probes, deliver CARS excitation pulses, and collect CARS signals. The adiabatic tapered fiber probe with a diameter of 11.61 µm can focus CARS excitation lights without mismatch at the focal point. The measurements for proof-of-concept were made with methanol, ethanol, cyclohexane, and acetone injected into simulated cells. The experimental results show that the tapered optical fiber probe can detect carbon-hydrogen (C–H) bond-rich substances and their concentration. To our best knowledge, this optical fiber probe provides the minimum size among probes for detecting CARS signals. These results pave the way for minimally invasive live-cell detection in the future.

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Fast epi-detected broadband multiplex CARS and SHG imaging of mouse skull cells

Cited by 22 publications

References 26 publications

Multiplex coherent anti-Stokes Raman scattering highlights state of chromatin condensation in CH region

Multiplex coherent anti-Stokes Raman scattering highlights state of chromatin condensation in CH region

Multi-modal Nonlinear Optical and Thermal Imaging Platform for Label-Free Characterization of Biological Tissue

Flexible Minimally Invasive CARS Measurement Method with Tapered Optical Fiber Probe for Single-Cell Application

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