Femtosecond stimulated Raman microscopy

Ploetz, Evelyn; Laimgruber, S.; Berner, Stefan; Zinth, Wolfgang; Gilch, Peter

doi:10.1007/s00340-007-2630-x

Cited by 302 publications

(229 citation statements)

References 24 publications

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“…The addition of Raman spectroscopy could be a rather simple but very beneficial step. By combining this with coherent Raman techniques, such as coherent anti-Stokes Raman scattering (CARS) [23,24,29,31] or stimulated Raman scattering (SRS) [30,32] even fast flow schemes analyzing thousands of cells will be possible, which is further aided by the spatial uniformity of Raman spectra in sperm cells. …”

Section: Discussionmentioning

confidence: 99%

“…Since then, the applications of Raman spectroscopy to single cells have grown exponentially [23,24], and applications related to single cancer cells [25], human embryonic stem cells [26], and even biological particles as small as lipoproteins [27] and bacterial spores [28] have been demonstrated. In addition, the field of coherent Raman microscopy has emerged, which enables label-free imaging of individual cells and tissues in vitro and in vivo [23,24,[29][30][31][32].…”

Section: Biophotonicsmentioning

confidence: 99%

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Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Huser

Orme

Hollars

et al. 2009

Journal of Biophotonics

110

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Healthy human males produce sperm cells of which about 25–40% have abnormal head shapes. Increases in the percentage of sperm exhibiting aberrant sperm head morphologies have been correlated with male infertility, and biochemical studies of pooled sperm have suggested that sperm with abnormal shape may contain DNA that has not been properly repackaged by protamine during spermatid development. We have used micro‐Raman spectroscopy to obtain Raman spectra from individual human sperm cells and examined how differences in the Raman spectra of sperm chromatin correlate with cell shape. We show that Raman spectra of individual sperm cells contain vibrational marker modes that can be used to assess the efficiency of DNA‐packaging for each cell. Raman spectra obtained from sperm cells with normal shape provide evidence that DNA in these sperm is very efficiently packaged. We find, however, that the relative protein content per cell and DNA packaging efficiencies are distributed over a relatively wide range for sperm cells with both normal and abnormal shape. These findings indicate that single cell Raman spectroscopy should be a valuable tool in assessing the quality of sperm cells for in‐vitro fertilization. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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Section: Discussionmentioning

confidence: 99%

Section: Biophotonicsmentioning

confidence: 99%

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Huser

Orme

Hollars

et al. 2009

Journal of Biophotonics

110

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“…Unfortunately, CARS suffers from spectral distortion, unwanted nonresonant background, nonstraightforward concentration dependence, and coherent image artifact (25). Most recently, SRS microscopy has emerged to supersede CARS microscopy in almost all aspects (29)(30)(31)(32)(33)(34)(35)(36)(37)(38). Using Einstein's stimulated emission principle (39,40), SRS has achieved unprecedented sensitivity down to ∼1,000 retinoic acid molecules and up to video rate imaging speed in vivo (30,33).…”

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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“…PLGA (700 mg) was dissolved in dichloromethane (5 ml) and atRA (7,35, or 70 mg) was added to the PLGA solution. Then, the mixture was dropped into 150 ml of aqueous solution (0.1 w/v% of PVA) using a glass syringe (10 ml) and a needle (needle gauge; 18 G) while stirring with a magnetic stirrer at 500 rpm.…”

Section: Preparation Of All Trans Retinoic Acid (Atra)-loaded Plga MImentioning

confidence: 99%

“…Several heterodyne detected optical imaging techniques based on interferometric CARS [1][2][3], stimulated Raman scattering (SRS) [4], transient absorption [5], and photothermal effect [6] have shown great potential for label-free imaging, e.g., SRS microscopy [7,[8][9][10][11] has been used for vibrational imaging of biomass [12], pharmaceutical samples [13,14], and lipid bodies [10,15]. All these techniques require the extraction of a small AC signal at the sub-microvolt level from a noisy environment.…”

Section: Introductionmentioning

confidence: 99%

Heterodyne detected nonlinear optical imaging in a lock‐in free manner

Slipchenko

Oglesbee

Zhang

et al. 2012

Journal of Biophotonics

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We report a compact, cost‐effective tuned amplifier for frequency‐selective amplification of the modulated signal in heterodyne detected nonlinear optical microscopy. Our method improved the signal to noise ratio by an order of magnitude compared to conventional lock‐in detection, as demonstrated through stimulated Raman scattering imaging of live cells and tissues at the speed of 2 μsec/pixel. Application of the tuned amplifier to transient absorption microscopy is also demonstrated. The increased signal to noise ratio allowed epi‐detected in vivo imaging of myelin and blood in rat spinal cord with high spatial resolution. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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Femtosecond stimulated Raman microscopy

Cited by 302 publications

References 24 publications

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

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

Heterodyne detected nonlinear optical imaging in a lock‐in free manner

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