In vivo Detection of Ferrous Cytochrome c in Mitochondria of Single Living Yeast Cells by Resonance Raman microspectroscopy

Onogi, Chikao; Hamaguchi, Hiro‐o; Champion, P. M.; Ziegler, L. D.

doi:10.1063/1.3482559

Cited by 6 publications

(9 citation statements)

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“…4,26 Raman microspectroscopy is considered as one of the newly established methods used for the detection of cytochrome c, since this molecule exhibits a characteristic vibrational band at 750 cm −1 and other bands in the mid-frequency spectral region (1131, 1312, 1585 cm −1 ); therefore this technique can be used to image cytochrome c without any labeling. [6][7][8][9] Raman spectra of pure cytochrome c from horse heart dissolved in PBS buffer at 250 μM have been recorded and results showed the expected vibrational bands (Fig. 6), in agreement with those published in literature.…”

Section: Resultssupporting

confidence: 89%

“…3 Mitochondria participate in apoptotic signaling by activating caspases via release of cytochrome c. Thus, detection of cytochrome c serves as a convenient marker for studying mitochondrial involvement in apoptosis. 4 Different spectroscopic approaches have so far been applied and developed to reach single molecule detection and particular attention has been focused on vibrational spectroscopies [5][6][7] because they are of fundamental importance for the understanding of internal configurations, structure and dynamics of proteins. In particular, Raman spectroscopy provides significantly detailed information on the structure and dynamics of molecules existing and playing roles in living cells.…”

Section: Introductionmentioning

confidence: 99%

“…8 Therefore, Raman spectroscopy is considered as one of the newly established methods used for the detection of cytochrome c, which shows a characteristic vibrational band at 750 cm −1 and other bands in the mid-frequency spectral region (1131, 1312, 1585 cm −1 ). [6][7][8][9] In the following study, dark toxicity and phototoxicity of aluminum phthalocyanine tetrasulfonate chloride were assessed. Confocal laser scanning microscopy was used to study the intracellular phthalocyanine localization and the morphological changes occurring upon photosensitization.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of photodynamic treatment using aluminum phthalocyanine tetrasulfonate chloride as a photosensitizer: new approach

Amin

Hauser²,

Kinzler³

et al. 2012

Photochem Photobiol Sci

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Photodynamic therapy (PDT) has been the subject of several clinical studies. Evidence to date suggests that direct cell death may involve apoptosis. T(24) cells (bladder cancer cells, ATCC-Nr. HTB-4) were subjected to PDT with aluminum phthalocyanine tetrasulfonate chloride (AlS(4)Pc-Cl) and red laser light at 670 nm. Morphological changes after PDT were visualized under confocal microscopy. Raman microspectroscopy is considered as one of the newly established methods used for the detection of cytochrome c as an apoptotic marker. Results showed that PDT treated T(24) cells seem to undergo apoptosis after irradiation with 3 J cm(-2). Cytochrome c could not be detected from cells incubated with AlS(4)Pc-Cl using Raman spectroscopy whereas AlS(4)Pc-Cl seems to interfere with the Raman spectrum of cytochrome c.

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Section: Resultssupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of photodynamic treatment using aluminum phthalocyanine tetrasulfonate chloride as a photosensitizer: new approach

Amin

Hauser²,

Kinzler³

et al. 2012

Photochem Photobiol Sci

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show abstract

“…In the past two decades, a number of publications have shown the successful application of Raman microspectroscopy to label-free molecular-level analysis of living cells and to discrimination of cell types. [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23] Although Raman spectra contain rich information on molecular structure, detailed interpretation of measured spectra is often difficult because of their complexity. Each Raman spectrum obtained from space-resolved mapping measurements is usually interpreted as a superposition of several spectral components of biomolecules, as well as a background and fluorescence.…”

Section: Introductionmentioning

confidence: 99%

Molecular component distribution imaging of living cells by multivariate curve resolution analysis of space-resolved Raman spectra

Ando

Hamaguchi

2013

J. Biomed. Opt

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Abstract. Label-free Raman microspectroscopy combined with a multivariate curve resolution (MCR) analysis can be a powerful tool for studying a wide range of biomedical molecular systems. The MCR with the alternating least squares (MCR-ALS) technique, which retrieves the pure component spectra from complicatedly overlapped spectra, has been successfully applied to in vivo and molecular-level analysis of living cells. The principles of the MCR-ALS analysis are reviewed with a model system of titanium oxide crystal polymorphs, followed by two examples of in vivo Raman imaging studies of living yeast cells, fission yeast, and budding yeast. Due to the non-negative matrix factorization algorithm used in the MCR-ALS analysis, the spectral information derived from this technique is just ready for physical and/or chemical interpretations. The corresponding concentration profiles provide the molecular component distribution images (MCDIs) that are vitally important for elucidating life at the molecular level, as stated by Schroedinger in his famous book, "What is life?" Without any a priori knowledge about spectral profiles, timeand space-resolved Raman measurements of a dividing fission yeast cell with the MCR-ALS elucidate the dynamic changes of major cellular components (lipids, proteins, and polysaccharides) during the cell cycle. The MCR-ALS technique also resolves broadly overlapped OH stretch Raman bands of water, clearly indicating the existence of organelle-specific water structures in a living budding yeast cell.

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“…The peak at ~1583 cm -1 was additionally identified in HeLa cells [52] and in yeast mitochondria. [53] The other spectral regions with notable differences in the Raman signal in Fig. 2 A and C for various excitations were observed at approximately 1437-1444 cm -1 (fatty acids, triglycerides, CH2 or CH3 deformation modes [54,55]) and ~2845-2940 cm -1 (CH2, CH3 stretching of lipids and proteins.…”

Section: Human Cancer Tissuementioning

confidence: 87%

Novel strategies of Raman imaging for monitoring intracellular retinoid metabolism in cancer cells

Abramczyk

Imiela

Surmacki

2020

Preprint

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We developed a label-free Raman method for whole-cell biochemical imaging to detect molecular processes that occur in normal and cancer brain cells due to retinol transport in human cancers at the level of isolated organelles. Our approach allows the creation of biochemical maps of lipid droplets, mitochondria and nuclei in single cells. The maps were capable of discriminating triglycerides (TAG) from retinyl ester (RE) in lipid droplets (LD), providing an excellent tool to monitor intracellular retinoid metabolism. We detected spectral changes that arose in proteins and lipids due to retinoid metabolism in human cell lines of normal astrocytes and high-grade cancer cells of glioblastoma as well as in human medulloblastoma and glioblastoma tissue. Raman imaging is an effective tool for monitoring of retinoids and retinol binding proteins involved in carcinogenesis, as monitored by the unique spectral signatures of vibrations. We found two functionally distinct lipid droplets: TAG-LD, for energy storage, and RE-LD, for regulating the level of apo-CRBP1 in cytosol. Raman polarization measurements revealed the occurrence of conformational changes affecting discrete regions of proteins associated with retinol binding. Aberrant expression of retinoids and retinol binding proteins in human tumours were localized in lipid droplets, mitochondria and nuclei according Raman imaging.

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In vivo Detection of Ferrous Cytochrome c in Mitochondria of Single Living Yeast Cells by Resonance Raman microspectroscopy

Cited by 6 publications

References 0 publications

Evaluation of photodynamic treatment using aluminum phthalocyanine tetrasulfonate chloride as a photosensitizer: new approach

Evaluation of photodynamic treatment using aluminum phthalocyanine tetrasulfonate chloride as a photosensitizer: new approach

Molecular component distribution imaging of living cells by multivariate curve resolution analysis of space-resolved Raman spectra

Novel strategies of Raman imaging for monitoring intracellular retinoid metabolism in cancer cells

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