In vivo multimode Raman imaging reveals concerted molecular composition and distribution changes during yeast cell cycle

Huang, Chih-Mao; Hamaguchi, Hiro O.; Shigeto, Shinsuke

doi:10.1039/c1cc12350e

Cited by 50 publications

(63 citation statements)

References 28 publications

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“…After wavelength calibration using the Raman spectrum of indene, all spectra acquired by the Raman mapping experiment were processed by a singular value decomposition analysis for noise reduction, as described in previous reports [19,36]. After the data pre-processing, for measurement of area intensity of the Raman band, the fitted spectrum was processed by gauss fitting method in the spectral region containing the Raman markers associated with cytochrome, protein, and AmB (726–762 cm −1 , 997–1009 cm −1 , 1533–1568 cm −1 , respectively).…”

Section: Methodsmentioning

confidence: 99%

In Situ Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman Microspectroscopy

et al. 2014

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The study of spatial distribution of secondary metabolites within microbial cells facilitates the screening of candidate strains from marine environments for functional metabolites and allows for the subsequent assessment of the production of metabolites, such as antibiotics. This paper demonstrates the first application of Raman microspectroscopy for in situ detection of the antifungal antibiotic amphotericin B (AmB) produced by actinomycetes—Streptomyces nodosus. Raman spectra measured from hyphae of S. nodosus show the specific Raman bands, caused by resonance enhancement, corresponding to the polyene chain of AmB. In addition, Raman microspectroscopy enabled us to monitor the time-dependent change of AmB production corresponding to the growth of mycelia. The Raman images of S. nodosus reveal the heterogeneous distribution of AmB within the mycelia and individual hyphae. Moreover, the molecular association state of AmB in the mycelia was directly identified by observed Raman spectral shifts. These findings suggest that Raman microspectroscopy could be used for in situ monitoring of antibiotic production directly in marine microorganisms with a method that is non-destructive and does not require labeling.

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Section: Methodsmentioning

confidence: 99%

In Situ Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman Microspectroscopy

et al. 2014

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“…In this work, we employed ultraviolet resonance Raman (UVRR) spectroscopy as a sensitive, specific reporter of drug‐induced changes in intact mammalian cells. In previous work, visible and near‐infrared Raman spectroscopies have been employed to monitor the viability of cells, study diseases and cellular events, and characterize drug‐biomolecular interactions ,. Compared to visible and near‐infrared Raman spectroscopy, UVRR spectroscopy generates less biofluorescence and produces more reliable baselines.…”

Section: Figurementioning

confidence: 99%

UV Resonance Raman Study of Apoptosis, Platinum‐Based Drugs, and Human Cell Lines

et al. 2018

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As a noninvasive molecular analysis technique, ultraviolet resonance Raman (UVRR) spectroscopy represents a label-free method suitable for characterizing biomolecules. Using UVRR spectroscopy, we collected spectral fingerprints of UV absorbing cellular components, including proteins, nucleic acids, and unsaturated lipids. This knowledge was used to guide the assignment of spectra derived from intact human cell lines (i. e., HSC-3 and HaCaT) and from the apoptotic events induced by cisplatin. Notably, a jet-flow system was employed to generate flowing cell suspensions during UVRR measurements, minimizing UV-induced damage. A spectral marker is established based on the ratio of Raman intensities at 1488 and 1655 cm ; this ratio correlates to the level of cell death due to apoptosis. Collectively, this work demonstrates that UVRR spectroscopy is a sensitive and informative probe of cellular physiology and molecular composition. The molecular insight obtained from UVRR measurements can be used to improve understanding of therapeutic treatment and to guide drug development and the choice of therapeutic agents.

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“…Those changes were correlated to cell cycle phase-to-phase transitions, i.e., G 1 -S and G 2 -M, and validated by flow cytometry. Huang and coworkers 50 investigated cell cycle-related changes in molecular composition, and molecular distribution in yeast cells in time-lapsed Raman-based imaging. Time-lapsed images based on Raman peaks associated with lipid and protein vibrations were used to show the distribution of biochemical components during the G 2 -M phase and G 1 -S phase.…”

Section: Biological Applicationsmentioning

confidence: 99%

Methods and Applications of Raman Microspectroscopy to Single-Cell Analysis

2013

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Raman spectroscopy is a powerful biochemical analysis technique that allows for the dynamic characterization and imaging of living biological cells in the absence of fluorescent stains. In this review, we summarize some of the most recent developments in the noninvasive biochemical characterization of single cells by spontaneous Raman scattering. Different instrumentation strategies utilizing confocal detection optics, multispot, and line illumination have been developed to improve the speed and sensitivity of the analysis of single cells by Raman spectroscopy. To analyze and visualize the large data sets obtained during such experiments, sophisticated multivariate statistical analysis tools are necessary to reduce the data and extract components of interest. We highlight the most recent applications of single cell analysis by Raman spectroscopy and their biomedical implications that have enabled the noninvasive characterization of specific metabolic states of eukaryotic cells, the identification and characterization of stem cells, and the rapid identification of bacterial cells. We conclude the article with a brief look into the future of this rapidly evolving research area.

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In vivo multimode Raman imaging reveals concerted molecular composition and distribution changes during yeast cell cycle

Abstract: In vivo time-lapse Raman imaging reveals highly dynamic and concerted changes in concentration and distribution of phospholipids and proteins during and after cell division of a single living Schizosaccharomyces pombe cell.

Cited by 50 publications

References 28 publications

In Situ Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman Microspectroscopy

In Situ Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman Microspectroscopy

UV Resonance Raman Study of Apoptosis, Platinum‐Based Drugs, and Human Cell Lines

Methods and Applications of Raman Microspectroscopy to Single-Cell Analysis

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