Label-free Raman observation of cytochrome c dynamics during apoptosis

Okada, Masaya; Smith, Nicholas I.; Palonpon, Almar F.; Endo, Hiromi; Kawata, Satoshi; Sodeoka, Mikiko; Fujita, Katsumasa

doi:10.1073/pnas.1107524108

Cited by 421 publications

(429 citation statements)

References 23 publications

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“…The most notable of these is heme, an iron-conjugated porphyrin which is found in hemoglobin in blood cells, and the metalloprotein cytochrome c. Heme also has an absorption band between 500-550 nm [98], which has allowed numerous studies by resonance Raman using 532 nm excitation, which produces spectra with strong bands, particularly the ring breathing mode of the pyrrole groups. Cytochrome c is a hemeprotein found in mitochondria, and its dynamics have been studied in apoptosing cells using resonance Raman of the 750 cm −1 pyrrole ring breathing band [113]. The oxygenation state of hemoglobin in single red blood cells can be also be observed with 785 nm-excited resonance Raman, in particular the Fe-O 2 stretching mode at 570 cm −1 was only seen for oxygenated cells [114].…”

Section: Heme and Other Natural Porphyrinsmentioning

confidence: 99%

Raman spectroscopy: techniques and applications in the life sciences

Shipp¹,

Sinjab²,

Notingher³

2017

Adv. Opt. Photon.

268

189

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Raman spectroscopy is an increasingly popular technique in many areas including biology and medicine.It is based on Raman scattering, a phenomenon in which incident photons lose or gain energy via interactions with vibrating molecules in a sample. These energy shifts can be used to obtain information regarding molecular composition of the sample with very high accuracy. Applications of Raman spectroscopy in the life sciences have included quantification of biomolecules, hyperspectral molecular imaging of cells and tissue, medical diagnosis, and others. This review briefly presents the physical origin of Raman scattering explaining the key classical and quantum mechanical concepts. Variations of the Raman effect will also be considered, including resonance, coherent, and enhanced Raman scattering. We discuss the molecular origins of prominent bands often found in the Raman spectra of biological samples. Finally, we examine several variations of Raman spectroscopy techniques in practice, looking at their applications, strengths, and challenges. This review is intended to be a starting resource for scientists new to Raman spectroscopy, providing theoretical background and practical examples as the foundation for further study and exploration.

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Section: Heme and Other Natural Porphyrinsmentioning

confidence: 99%

Raman spectroscopy: techniques and applications in the life sciences

Shipp¹,

Sinjab²,

Notingher³

2017

Adv. Opt. Photon.

268

189

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“…[4][5][6][7][8] RMS was also used for time-course experiments on individual live cells revealing molecular processes not attainable with other imaging. [9][10][11][12][13] The integration of environmental chambers with inverted Raman microscopes can allow time-and spatially-resolved molecular studies of cellular processes spanning days and weeks, such as stem cell differentiation. 14,15 Nevertheless, RMS has been used only in few studies to characterize molecular details associated with host-parasite interaction.…”

Section: Introductionmentioning

confidence: 99%

Analysis of interaction between the apicomplexan protozoan Toxoplasma gondii and host cells using label-free Raman spectroscopy

Naemat

Elsheikha

Al-sandaqchi

et al. 2015

Analyst

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Label-free imaging using Raman micro-spectroscopy (RMS) was used to characterize the spatio-temporal molecular changes of T. gondii tachyzoites and their host cell microenvironment. Raman spectral maps were recorded from isolated T. gondii tachyzoites and T. gondii-infected human retinal cells at 6 hr, 24 hr and 48 hr postinfection. Principal component analysis (PCA) of the Raman spectra of paraformaldehyde-fixed infected cells indicated a significant increase in the amount of lipids and proteins in the T. gondii tachyzoites as the infection progresses within host cells. These results were confirmed by experiments carried out on live T. gondii-infected cells and were correlated with an increase in the concentration of proteins and lipids required for the replication of this intracellular pathogen. These findings demonstrate the potential of RMS to characterize time-and spatially-dependent molecular interactions between intracellular pathogens and the host cells. Such information may be useful for discovery of pharmacological targets or screening compounds with potential neuroprotective activity for eminent effects of changes in brain infection control practices. 2

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“…Nevertheless, it has unique capabilities, including the observation of living organisms in aqueous and physiological environments (14,15). In addition, intrinsic vibrational modes of membrane lipids in Raman spectra can provide a great deal of molecular information about the composition, structure, and phase state of lipid molecules (12).…”

Section: Discussionmentioning

confidence: 99%

Sphingomyelin distribution in lipid rafts of artificial monolayer membranes visualized by Raman microscopy

Ando

Kinoshita

Cui

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

125

102

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Sphingomyelin (SM) and cholesterol (chol)-rich domains in cell membranes, called lipid rafts, are thought to have important biological functions related to membrane signaling and protein trafficking. To visualize the distribution of SM in lipid rafts by means of Raman microscopy, we designed and synthesized an SM analog tagged with a Raman-active diyne moiety (diyne-SM). Diyne-SM showed a strong peak in a Raman silent region that is free of interference from intrinsic vibrational modes of lipids and did not appear to alter the properties of SM-containing monolayers. Therefore, we used Raman microscopy to directly visualize the distribution of diyne-SM in raft-mimicking domains formed in SM/dioleoylphosphatidylcholine/chol ternary monolayers. Raman images visualized a heterogeneous distribution of diyne-SM, which showed marked variation, even within a single ordered domain. Specifically, diyne-SM was enriched in the central area of raft domains compared with the peripheral area. These results seem incompatible with the generally accepted raft model, in which the raft and nonraft phases show a clear biphasic separation. One of the possible reasons is that gradual changes of SM concentration occur between SM-rich and -poor regions to minimize hydrophobic mismatch. We believe that our technique of hyperspectral Raman imaging of a single lipid monolayer opens the door to quantitative analysis of lipid membranes by providing both chemical information and spatial distribution with high (diffraction-limited) spatial resolution.lipid raft | Raman imaging | alkyne tag | supported monolayer | sphingomyelin

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Label-free Raman observation of cytochrome c dynamics during apoptosis

Cited by 421 publications

References 23 publications

Raman spectroscopy: techniques and applications in the life sciences

Raman spectroscopy: techniques and applications in the life sciences

Analysis of interaction between the apicomplexan protozoan Toxoplasma gondii and host cells using label-free Raman spectroscopy

Sphingomyelin distribution in lipid rafts of artificial monolayer membranes visualized by Raman microscopy

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