9,688 research outputs found
Quantitative volumetric Raman imaging of three dimensional cell cultures
The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in 3D cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy
Coherent Raman spectro-imaging with laser frequency combs
Optical spectroscopy and imaging of microscopic samples have opened up a wide
range of applications throughout the physical, chemical, and biological
sciences. High chemical specificity may be achieved by directly interrogating
the fundamental or low-lying vibrational energy levels of the compound
molecules. Amongst the available prevailing label-free techniques, coherent
Raman scattering has the distinguishing features of high spatial resolution
down to 200 nm and three-dimensional sectioning. However, combining fast
imaging speed and identification of multiple - and possibly unexpected-
compounds remains challenging: existing high spectral resolution schemes
require long measurement times to achieve broad spectral spans. Here we
overcome this difficulty and introduce a novel concept of coherent anti-Stokes
Raman scattering (CARS) spectro-imaging with two laser frequency combs. We
illustrate the power of our technique with high resolution (4 cm-1) Raman
spectra spanning more than 1200 cm-1 recorded within less than 15 microseconds.
Furthermore, hyperspectral images combining high spectral (10 cm-1) and spatial
(2 micrometers) resolutions are acquired at a rate of 50 pixels per second.
Real-time multiplex accessing of hyperspectral images may dramatically expand
the range of applications of nonlinear microscopy.Comment: 8 pages, 3 figure
Seafloor characterization using airborne hyperspectral co-registration procedures independent from attitude and positioning sensors
The advance of remote-sensing technology and data-storage capabilities has progressed in the last decade to commercial multi-sensor data collection. There is a constant need to characterize, quantify and monitor the coastal areas for habitat research and coastal management. In this paper, we present work on seafloor characterization that uses hyperspectral imagery (HSI). The HSI data allows the operator to extend seafloor characterization from multibeam backscatter towards land and thus creates a seamless ocean-to-land characterization of the littoral zone
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Pre-processing visualization of hyperspectral fluorescent data with Spectrally Encoded Enhanced Representations.
Hyperspectral fluorescence imaging is gaining popularity for it enables multiplexing of spatio-temporal dynamics across scales for molecules, cells and tissues with multiple fluorescent labels. This is made possible by adding the dimension of wavelength to the dataset. The resulting datasets are high in information density and often require lengthy analyses to separate the overlapping fluorescent spectra. Understanding and visualizing these large multi-dimensional datasets during acquisition and pre-processing can be challenging. Here we present Spectrally Encoded Enhanced Representations (SEER), an approach for improved and computationally efficient simultaneous color visualization of multiple spectral components of hyperspectral fluorescence images. Exploiting the mathematical properties of the phasor method, we transform the wavelength space into information-rich color maps for RGB display visualization. We present multiple biological fluorescent samples and highlight SEER's enhancement of specific and subtle spectral differences, providing a fast, intuitive and mathematical way to interpret hyperspectral images during collection, pre-processing and analysis
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