The need for speed

Suhalim, Jeffrey L.; Boik, John C.; Tromberg, Bruce J.; Potma, Eric O.

doi:10.1002/jbio.201200002

Cited by 59 publications

(42 citation statements)

References 64 publications

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“…Raman signals can be enhanced in tissue or cells by surface enhanced Raman scattering (SERS) effect using functionalized metal nanoparticles [105][106][107] or by resonance Raman scattering probing prosthetic groups in chromophores [108]. Another promising signal enhancement approach constitutes non-linear, coherent Raman techniques such as coherent anti-Stokes Raman scattering (CARS) or stimulated Raman scattering [109]. CARS imaging offers video rate acquisition speed and enables detection of single cells and nuclei probing the CHstretching wavenumber range [110].…”

Section: Resultsmentioning

confidence: 99%

Molecular pathology via IR and Raman spectral imaging

Diem¹,

Mazur²,

Lenau³

et al. 2013

Journal of Biophotonics

177

115

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Journal of BIOPHOTONICSDuring the last 15 years, vibrational spectroscopic methods have been developed that can be viewed as molecular pathology methods that depend on sampling the entire genome, proteome and metabolome of cells and tissues, rather than probing for the presence of selected markers. First, this review introduces the background and fundamentals of the spectroscopies underlying the new methodologies, namely infrared and Raman spectroscopy. Then, results are presented in the context of spectral histopathology of tissues for detection of metastases in lymph nodes, squamous cell carcinoma, adenocarcinomas, brain tumors and brain metastases. Results from spectral cytopathology of cells are discussed for screening of oral and cervical mucosa, and circulating tumor cells. It is concluded that infrared and Raman spectroscopy can complement histopathology and reveal information that is available in classical methods only by costly and time-consuming steps such as immunohistochemistry, polymerase chain reaction or gene arrays. Due to the inherent sensitivity toward changes in the bio-molecular composition of different cell and tissue types, vibrational spectroscopy can even provide information that is in some cases superior to that of any one of the conventional techniques.Schematic of spectral histopathology (SHP) process: diagnostic algorithm is developed by spectral data of well documented specimens and subsequently applied to unknown dataset.

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Section: Resultsmentioning

confidence: 99%

Molecular pathology via IR and Raman spectral imaging

Diem¹,

Mazur²,

Lenau³

et al. 2013

Journal of Biophotonics

177

115

View full text Add to dashboard Cite

show abstract

“…This offers a way to investigate the orientation of the collagen fibrils [67,83]. Given that the amide I peak characterizes the organic and the v 1 phosphate peak the inorganic part of the ultrastructure, polarized Raman spectroscopy (PRS) can be used to derive information on the collagen and Except if coherent Raman scattering is used [61], which in exchange leads to significantly higher costs.…”

Section: Polarized Raman Spectroscopymentioning

confidence: 99%

“…In addition, Raman spectroscopy is performed in reflection mode, meaning that it can be used to analyse a sample without the need to section it, even in vivo [87], and can reach the bone under the skin [88,89]. On the other hand, Raman experiments are much more time-consuming (acquisition of one spectrum typically needs tens of seconds, except if coherent Raman scattering is employed [61]), and offer a lower spatial resolution of approximately 1 mm compared with that of PLM (approx. 250 nm).…”

Section: Polarized Raman Spectroscopymentioning

confidence: 99%

Techniques to assess bone ultrastructure organization: orientation and arrangement of mineralized collagen fibrils

Georgiadis

Müller

Schneider

2016

J. R. Soc. Interface.

113

100

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Bone's remarkable mechanical properties are a result of its hierarchical structure. The mineralized collagen fibrils, made up of collagen fibrils and crystal platelets, are bone's building blocks at an ultrastructural level. The organization of bone's ultrastructure with respect to the orientation and arrangement of mineralized collagen fibrils has been the matter of numerous studies based on a variety of imaging techniques in the past decades. These techniques either exploit physical principles, such as polarization, diffraction or scattering to examine bone ultrastructure orientation and arrangement, or directly image the fibrils at the sub-micrometre scale. They make use of diverse probes such as visible light, X-rays and electrons at different scales, from centimetres down to nanometres. They allow imaging of bone sections or surfaces in two dimensions or investigating bone tissue truly in three dimensions, in vivo or ex vivo, and sometimes in combination with in situ mechanical experiments. The purpose of this review is to summarize and discuss this broad range of imaging techniques and the different modalities of their use, in order to discuss their advantages and limitations for the assessment of bone ultrastructure organization with respect to the orientation and arrangement of mineralized collagen fibrils.

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“…However, spontaneous Raman scattering is an intrinsically weak process, hence not ideal for fast live-cell imaging (23). As a nonlinear technique, coherent antiStokes Raman scattering (CARS) offers much higher imaging speed by virtue of coherent amplification (24)(25)(26)(27)(28). Unfortunately, CARS suffers from spectral distortion, unwanted nonresonant background, nonstraightforward concentration dependence, and coherent image artifact (25).…”

mentioning

confidence: 99%

Vibrational imaging of newly synthesized proteins in live cells by stimulated Raman scattering microscopy

Lu¹,

Shen³

et al. 2013

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

211

223

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Synthesis of new proteins, a key step in the central dogma of molecular biology, has been a major biological process by which cells respond rapidly to environmental cues in both physiological and pathological conditions. However, the selective visualization of a newly synthesized proteome in living systems with subcellular resolution has proven to be rather challenging, despite the extensive efforts along the lines of fluorescence staining, autoradiography, and mass spectrometry. Herein, we report an imaging technique to visualize nascent proteins by harnessing the emerging stimulated Raman scattering (SRS) microscopy coupled with metabolic incorporation of deuterium-labeled amino acids. As a first demonstration, we imaged newly synthesized proteins in live mammalian cells with high spatial-temporal resolution without fixation or staining. Subcellular compartments with fast protein turnover in HeLa and HEK293T cells, and newly grown neurites in differentiating neuron-like N2A cells, are clearly identified via this imaging technique. Technically, incorporation of deuterium-labeled amino acids is minimally perturbative to live cells, whereas SRS imaging of exogenous carbon-deuterium bonds (C-D) in the cell-silent Raman region is highly sensitive, specific, and compatible with living systems. Moreover, coupled with label-free SRS imaging of the total proteome, our method can readily generate spatial maps of the quantitative ratio between new and total proteomes. Thus, this technique of nonlinear vibrational imaging of stable isotope incorporation will be a valuable tool to advance our understanding of the complex spatial and temporal dynamics of newly synthesized proteome in vivo.stable isotope labeling | stimulated Raman microscopy | protein synthesis

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The need for speed

Cited by 59 publications

References 64 publications

Molecular pathology via IR and Raman spectral imaging

Molecular pathology via IR and Raman spectral imaging

Techniques to assess bone ultrastructure organization: orientation and arrangement of mineralized collagen fibrils

Vibrational imaging of newly synthesized proteins in live cells by stimulated Raman scattering microscopy

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