Raman microspectroscopic approach to the study of human granulocytes

Puppels, Gerwin J.; Garritsen, Henk; Segers-Nolten, Gezina M.J.; Mul, F.F.M. de; Greve, Jan

doi:10.1016/s0006-3495(91)82142-7

Cited by 171 publications

(156 citation statements)

References 46 publications

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“…The Raman spectrum from the green cluster (Fig. 1C, spectrum 2) is typical for the cytoplasm (32) and differs from the red cluster (spectrum not shown) mainly in its higher overall intensity. Because LBs in cells are easily recognized by Raman spectroscopy because their high aliphatic chain density gives rise to very strong Raman signals (33), the absence of a cluster containing strong lipid signals in the image shown in Fig.…”

Section: Resultsmentioning

confidence: 95%

Single-cell Raman and fluorescence microscopy reveal the association of lipid bodies with phagosomes in leukocytes

Manen

Kraan²,

Roos

et al. 2005

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

288

247

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Cellular imaging techniques based on vibrational spectroscopy have become powerful tools in cell biology because the molecular composition of subcellular compartments can be visualized without the need for labeling. Using high-resolution, nonresonant confocal Raman microscopy on individual cells, we demonstrate here that lipid bodies (LBs) rich in arachidonate as revealed by their Raman spectra associate with latex bead-containing phagosomes in neutrophilic granulocytes. This finding was corroborated in macrophages and in PLB-985 cells, which can be induced to differentiate into neutrophil-like cells, by selective staining of LBs and visualization by confocal fluorescence microscopy. We further show that the accumulation of LBs near phagosomes is mediated at least in part by the flavohemoprotein gp91 phox (in which ''phox'' is phagocyte oxidase), because different LB distributions around phagocytosed latex beads were observed in WT and gp91 phoxdeficient PLB-985 cells. gp91 phox , which accumulates in the phagosomal membrane, is the catalytic subunit of the leukocyte NADPH oxidase, a critical enzyme in the innate immune response. Finally, time-lapse fluorescence microscopy experiments on neutrophils revealed that the LB-phagosome association is transient, similar to the ''kiss-and-run'' behavior displayed by endosomes involved in phagosome maturation. Because arachidonic acid (AA) has been shown to be involved in NADPH oxidase activation and phagosome maturation in neutrophils and macrophages, respectively, the findings reported here suggest that LBs may provide a reservoir of AA for local activation of these essential leukocyte functions.innate immunity ͉ NADPH oxidase ͉ Raman microscopy ͉ phagocytes

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Section: Resultsmentioning

confidence: 95%

Single-cell Raman and fluorescence microscopy reveal the association of lipid bodies with phagosomes in leukocytes

Manen

Kraan²,

Roos

et al. 2005

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

288

247

View full text Add to dashboard Cite

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“…The specific information contained in the Raman spectra can be related to changes to the physiology as a result of external stimuli [27][28][29][30]. The spatial resolution is of the order of 0.5-2 mm, providing access to the subcellular organisation of the cells at a molecular level [31][32][33][34][35]. In the case of FTIR spectroscopy, the spatial resolution is limited to some 5-10 mm in free space applications, although this can be reduced to 1-2 mm using high numerical aperture microscopic objectives, as in the case of Attenuated Total Reflection (ATR) FTIR microspectroscopy.…”

Section: Biophotonicsmentioning

confidence: 99%

Improved protocols for vibrational spectroscopic analysis of body fluids

Bonnier¹,

Petitjean

Baker

et al. 2013

Journal of Biophotonics

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The applications of vibrational spectroscopy to the examination of human blood serum are explored. Although FTIR spectra can be recorded in aqueous solutions at (gelatin) concentrations as low as 100 mg/L, the high-wavenumber region remains obscured by water absorption. Using Raman spectroscopy, high quality spectra of gelatine solutions as low as 10 mg/L can be achieved, also covering the high-wavenumber regions. In human serum, spectral profiles are weak and partially obscured by water features. Dried deposits are shown to be physically and chemically inhomogeneous resulting in reduced measurement reproducibility. Concentration of the serum using commercially available centrifugal filter devices results in an improvement in the spectral intensity and quality. Additionally, in Raman spectroscopy, reduced background and significantly enhanced signal collection is achievable by measurement in an inverted geometry. The improved protocols for spectroscopic measurement of human serum are applicable to a range of bodily fluids and should accelerate potential clinical applications

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“…Other important developments accelerating the progress of Raman spectroscopy include the digitization of spectra using charge-coupled devices (CCDs) [7,8], the confocal Raman microscope [9], and improved filters to remove light at the laser wavelength [10]. These inventions allowed a rapid increase in the popularity of using Raman to study biological samples in the early 1990s [11][12][13].…”

Section: Introductionmentioning

confidence: 99%

Raman spectroscopy: techniques and applications in the life sciences

Shipp¹,

Sinjab²,

Notingher³

2017

Adv. Opt. Photon.

255

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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Raman microspectroscopic approach to the study of human granulocytes

Cited by 171 publications

References 46 publications

Single-cell Raman and fluorescence microscopy reveal the association of lipid bodies with phagosomes in leukocytes

Single-cell Raman and fluorescence microscopy reveal the association of lipid bodies with phagosomes in leukocytes

Improved protocols for vibrational spectroscopic analysis of body fluids

Raman spectroscopy: techniques and applications in the life sciences

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