Separation of CARS image contributions with a Gaussian mixture model

Vogler, Nadine; Bocklitz, Thomas; Mariani, Melissa M.; Deckert, Volker; Marková, Aneta; Schelkens, Peter; Rösch, Petra; Akimov, D. A.; Dietzek, Benjamin; Popp, Jürgen

doi:10.1364/josaa.27.001361

Cited by 22 publications

(17 citation statements)

References 43 publications

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“…Furthermore, statistic image analysis can be applied to interpret CARS images. 46,47 These first results are very promising. A combination of nonlinear imaging techniques visualized the morphology of brain tissue and differentiated cancerous and necrotic brain tissue.…”

Section: Discussionmentioning

confidence: 91%

Nonlinear microscopy, infrared, and Raman microspectroscopy for brain tumor analysis

et al. 2011

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Contemporary brain tumor research focuses on two challenges: First, tumor typing and grading by analyzing excised tissue is of utmost importance for choosing a therapy. Second, for prognostication the tumor has to be removed as completely as possible. Nowadays, histopathology of excised tissue using haematoxylin-eosine staining is the gold standard for the definitive diagnosis of surgical pathology specimens. However, it is neither applicable in vivo, nor does it allow for precise tumor typing in those cases when only nonrepresentative specimens are procured. Infrared and Raman spectroscopy allow for very precise cancer analysis due to their molecular specificity, while nonlinear microscopy is a suitable tool for rapid imaging of large tissue sections. Here, unstained samples from the brain of a domestic pig have been investigated by a multimodal nonlinear imaging approach combining coherent anti-Stokes Raman scattering, second harmonic generation, and two photon excited fluorescence microscopy. Furthermore, a brain tumor specimen was additionally analyzed by linear Raman and Fourier transform infrared imaging for a detailed assessment of the tissue types that is required for classification and to validate the multimodal imaging approach. Hence label-free vibrational microspectroscopic imaging is a promising tool for fast and precise in vivo diagnostics of brain tumors.

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Section: Discussionmentioning

confidence: 91%

Nonlinear microscopy, infrared, and Raman microspectroscopy for brain tumor analysis

et al. 2011

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“…4, 5, and 10 was described previously ( 25 ). For the experiments, narrowband (bandwidth, 0.5-1 nm) ps pulses were used to match the C-H stretching region at about 2,900 cm Ϫ 1 .…”

Section: Cars Microscopymentioning

confidence: 99%

Fluorescence-based fixative and vital staining of lipid droplets in Caenorhabditis elegans reveal fat stores using microscopy and flow cytometry approaches

Klapper

Ehmke

Palgunow

et al. 2011

Journal of Lipid Research

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Body composition with respect to white adipose tissue and lean body mass is of central importance and a determining factor of adiposity-associated perturbations, such as diabetes type 2, cardiovascular disease, and cancer. Therefore, modulation of body composition and especially the reduction of body fat are the main foci of dietary and lifestyle interventions. In the last few years, Caenorhabditis elegans became a model organism for investigating fat storage and metabolism, as reviewed by different authors ( 1-3 ). Based on forward genetics ( 4-6 ), functional genomics, and candidate gene approaches ( 7,8 ), more than 400 genes possibly involved in fat storage were described in C. elegans . Most of these genes are evolutionarily conserved and allocated in common pathways and are of relevance for the study of human adiposity.Although C. elegans was extensively used as a model organism to study the genetic and functional basis of fat storage, robust lipid droplet markers such as perilipin are not available at the moment ( 2 ). Appropriate methods concerning the distribution behavior of dyes in the worm or the reproducibility of staining procedures are still under discussion ( 3 ). The classical fi xative Sudan black dye ( 9 ) is shown to be very error prone in C. elegans because of the last ethanol-based washing steps. Recently O' Rourke et al. ( 10 ) demonstrated that the vital dyes Nile red and C1-C12-BODIPY-labeled fatty acid, which were used in more than 75 reports, stain lysosome-like granules rather than lipid droplets within the C. elegans intestine. These dyes were shown not to localize with the fi xative neutral fat fl uorophore LipidTox ( 10 ) 17 March 2011. Published, JLR Papers in Press, March 18, 2011 DOI 10.1194 Fluorescence-based fi xative and vital staining of lipid droplets in Caenorhabditis elegans reveal fat stores using microscopy and fl ow cytometry approaches Abbreviations: BP, bandpass; CARS, coherent anti-Stokes Raman scattering; COPAS, cytometry-based object parametric analysis and sorting system; LRO, lysosome-related organelle; OPO, optical parametric oscillator; TOF, time of fl ight; TAG, triacylglycerol. Manuscript received 8 October 2010 and in revised form

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“…8 Thus, the use of picoseconds pulses where the bandwidth of the lasers matches the spectral width of the vibrational transitions is a straight forward approach to keep the background as low as possible or to optimize the ratio between resonant signal (baring chemical contrast) versus nonresonant signal. 8 Overall, several approaches 7 have been suggested to reduce the impact of this nonlinear background and contrast reducing nonresonant CARS signals such as heterodyne CARS (i.e., interferometric mixing), 27,28 polarization sensitive CARS, 29 frequency modulation CARS, 30 application of image-processing approaches, 31 or the application of EPI-CARS to detect exclusively small objects of a size determined by the phase mismatch Δk (see also further above). 22,23 It needs to be kept in mind that the quality of single-band CARS images depends on: (1) the Raman scattering cross section of the mode to be imaged, (2) that this mode is spectrally distinct, i.e., can be found in a wavenumber silent region to avoid spectral interferences with adjacent Raman modes, and (3) on a high enough molecular density of oscillators featuring this mode.…”

Section: Single-band Cars Microscopymentioning

confidence: 99%

Raman and coherent anti-Stokes Raman scattering microspectroscopy for biomedical applications

et al. 2012

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Abstract. A tutorial article is presented for the use of linear and nonlinear Raman microspectroscopies in biomedical diagnostics. Coherent anti-Stokes Raman scattering (CARS) is the most frequently applied nonlinear variant of Raman spectroscopy. The basic concepts of Raman and CARS are introduced first, and subsequent biomedical applications of Raman and CARS are described. Raman microspectroscopy is applied to both in-vivo and in-vitro tissue diagnostics, and the characterization and identification of individual mammalian cells. These applications benefit from the fact that Raman spectra provide specific information on the chemical composition and molecular structure in a label-free and nondestructive manner. Combining the chemical specificity of Raman spectroscopy with the spatial resolution of an optical microscope allows recording hyperspectral images with molecular contrast. We also elaborate on interfacing Raman spectroscopic tools with other technologies such as optical tweezing, microfluidics and fiber optic probes. Thereby, we aim at presenting a guide into one exciting branch of modern biophotonics research.

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Separation of CARS image contributions with a Gaussian mixture model

Cited by 22 publications

References 43 publications

Nonlinear microscopy, infrared, and Raman microspectroscopy for brain tumor analysis

Nonlinear microscopy, infrared, and Raman microspectroscopy for brain tumor analysis

Fluorescence-based fixative and vital staining of lipid droplets in Caenorhabditis elegans reveal fat stores using microscopy and flow cytometry approaches

Raman and coherent anti-Stokes Raman scattering microspectroscopy for biomedical applications

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