Wide Field Spectral Imaging with Shifted Excitation Raman Difference Spectroscopy Using the Nod and Shuffle Technique

Wide field Raman imaging using the integral field spectroscopy approach was used as a fast, one shot imaging method for the simultaneous collection of all spectra composing a Raman image. For the suppression of autofluorescence and background signals such as room light, shifted excitation Raman difference spectroscopy (SERDS) was applied to remove background artifacts in Raman spectra. To reduce acquisition times in wide field SERDS imaging, we adapted the nod and shuffle technique from astrophysics and implemented it into a wide field SERDS imaging setup. In our adapted version, the nod corresponds to the change in excitation wavelength, whereas the shuffle corresponds to the shifting of charges up and down on a Charge-Coupled Device (CCD) chip synchronous to the change in excitation wavelength. We coupled this improved wide field SERDS imaging setup to diode lasers with 784.4/785.5 and 457.7/458.9 nm excitation and applied it to samples such as paracetamol and aspirin tablets, polystyrene and polymethyl methacrylate beads, as well as pork meat using multiple accumulations with acquisition times in the range of 50 to 200 ms. The results tackle two main challenges of SERDS imaging: gradual photobleaching changes the autofluorescence background, and multiple readouts of CCD detector prolong the acquisition time.Comment: Accepted and Published by "Sensors" Journal, 19 pages, 8 figure

Rotational CARS application to simultaneous and multiple-point temperature and concentration determination in a turbulent flow

Coherent anti-Stokes Raman scattering (CARS) from the pure rotational Raman lines of N2 is employed to measure the instantaneous (approximately 10 ns) rotational temperature of N2 gas at room temperature and below with good spatial resolution (0.2 x 0.2 x 3.0 cu mm). A broad bandwidth dye laser is used to obtain the entire rotational spectrum from a single laser pulse; the CARS signal is then dispersed by a spectrograph and recorded on an optical multichannel analyzer. A best fit temperature is found in several seconds with the aid of a computer for each experimental spectrum by a least squares comparison with calculated spectra. The model used to calculate the theoretical spectra incorporates the temperature and pressure dependence of the pressure-broadened rotational Raman lines, includes the nonresonant background susceptibility, and assumes that the pump laser has a finite linewidth. Temperatures are fit to experimental spectra recorded over the temperature range of 135 to 296 K, and over the pressure range of .13 to 15.3 atm

Wide Field Spectral Imaging with Shifted Excitation Raman Difference Spectroscopy Using the Nod and Shuffle Technique

Wide field Raman imaging using the integral field spectroscopy approach was used as a fast, one shot imaging method for the simultaneous collection of all spectra composing a Raman image. For the suppression of autofluorescence and background signals such as room light, shifted excitation Raman difference spectroscopy (SERDS) was applied to remove background artifacts in Raman spectra. To reduce acquisition times in wide field SERDS imaging, we adapted the nod and shuffle technique from astrophysics and implemented it into a wide field SERDS imaging setup. In our adapted version, the nod corresponds to the change in excitation wavelength, whereas the shuffle corresponds to the shifting of charges up and down on a Charge-Coupled Device (CCD) chip synchronous to the change in excitation wavelength. We coupled this improved wide field SERDS imaging setup to diode lasers with 784.4/785.5 and 457.7/458.9 nm excitation and applied it to samples such as paracetamol and aspirin tablets, polystyrene and polymethyl methacrylate beads, as well as pork meat using multiple accumulations with acquisition times in the range of 50 to 200 ms. The results tackle two main challenges of SERDS imaging: gradual photobleaching changes the autofluorescence background, and multiple readouts of CCD detector prolong the acquisition time

Multichannel Diffuse Optical Raman Tomography for Bone Characterization In Vivo: a Phantom Study

Raman spectroscopy is used to gather information on the mineral and organic components of bone tissue to analyze their composition. By measuring the Raman signal of bone through spatially offset Raman spectroscopy the health of the bone can be determined. We’ve customized a system with 8 collection channels that consist of individual fibers, which are coupled to separate spectrometers and cooled CCDs. This parallel detection system was used to scan gelatin phantoms with Teflon inclusions of two sizes. Raman signals were decoupled from the autofluorescence background using channel specific polynomial fitting. Images with high contrast to background ratios of Raman yield and accurate spatial resolution were recovered using a model-based diffuse tomography approach

Ultrafast imaging Raman spectroscopy of large-area samples without stepwise scanning

Step-by-step, time-consuming scanning of the sample is still the state-of-the-art in imaging Raman spectroscopy. Even for a few 100 image points the measurement time may add up to minutes or hours. A radical decrease in measurement time can be achieved by applying multiplex spectrographs coupled to imaging fiber bundles that are successfully used in astronomy. For optimal use of the scarce and expensive observation time at astronomical observatories, special high-performance spectrograph systems were developed. They are designed for recording thousands of spatially resolved spectra of a two-dimensional image field within one single exposure. Transferring this technology to imaging Raman spectroscopy allows a considerably faster acquisition of chemical maps. Currently, an imaging field of up to 1 cm2 can be investigated. For porcine skin the required measurement time is less than 1 min. For this reason, this technique is of particular interest for medical diagnostics, e.g., the identification of potentially cancerous abnormalities of skin tissue

The development of biomolecular Raman optical activity spectroscopy

Following its first observation over 40 years ago, Raman optical activity (ROA), which may be measured as a small difference in the intensity of vibrational Raman scattering from chiral molecules in right- and left-circularly polarized incident light or, equivalently, the intensity of a small circularly polarized component in the scattered light using incident light of fixed polarization, has evolved into a powerful chiroptical spectroscopy for studying a large range of biomolecules in aqueous solution. The long and tortuous path leading to the first observations of ROA in biomolecules in 1989, in which the author was closely involved from the very beginning, is documented, followed by a survey of subsequent developments and applications up to the present day. Among other things, ROA provides information about motif and fold, as well as secondary structure, of proteins; solution structure of carbohydrates; polypeptide and carbohydrate structure of intact glycoproteins; new insight into structural elements present in unfolded protein sequences; and protein and nucleic acid structure of intact viruses. Quantum chemical simulations of observed Raman optical activity spectra provide the complete three-dimensional structure, together with information about conformational dynamics, of smaller biomolecules. Biomolecular ROA measurements are now routine thanks to a commercial instrument based on a novel design becoming available in 2004

Identification of microplastics using Raman spectroscopy: latest developments and future prospects

Widespread microplastic pollution is raising growing concerns as to its detrimental effects upon living organisms. A realistic risk assessment must stand on representative data on the abundance, size distribution and chemical composition of microplastics. Raman microscopy is an indispensable tool for the analysis of very small microplastics (<20 μm). Still, its use is far from widespread, in part due to drawbacks such as long measurement time and proneness to spectral distortion induced by fluorescence. This review discusses each drawback followed by a showcase of interesting and easily available solutions that contribute to faster and better identification of microplastics using Raman spectroscopy. Among discussed topics are: enhanced signal quality with better detectors and spectrum processing; automated particle selection for faster Raman mapping; comprehensive reference libraries for successful spectral matching. A last section introduces non-conventional Raman techniques (non-linear Raman, hyperspectral imaging, standoff Raman) which permit more advanced applications such as real-time Raman detection and imaging of microplastics.publishe

Innovative approaches to selective detection and remote analysis : developments in surface-enhanced raman scattering (sers)-based and separations-based fiberoptic chemical sensors

This two-part study investigates the feasibility of selective detection in remote analysis based on 1) surface-enhanced Raman scattering (SERS), and 2) separations-based fiberoptic sensing (SBFOS). For the first case, the extremely sharp spectral features of Raman scattering can sometimes allow the analysis of multicomponent samples without complicated sample pretreatment steps. Furthermore, the giant signal-enhancing effect of SERS can enable trace level detection. A solid, surface-based metallic substrate approach is taken for the development of a practical SERS technology. Various substrates are described, including silver-coated alumina, silver-coated Ti02, and silver islands. These substrates are economical and easy to fabricate with a high degree of reproducibility. Furthermore, they are readily integrated with fiberoptic sensors for remote SERS detection. Three fiberoptic SERS sensor systems are described in this work. The surface-based substrates are also applied to the detection of organic vapors. In the latter case, the high separation efficiency of capillary electrophoresis is coupled with laser-induced fluorescence (LIF) detection in the development of a fiberoptic sensor. Although LIF can offer exceptional detectability, its application to the analysis of complex samples can be difficult due to the broadband nature of fluorescence. It often requires sample pretreatment steps such as separations. In the SBFOS approach, separations can be performed remotely. Several complications are associated with the development of CE-based SBFOSs and are described in this work. Four SBFOS designs are described and applied to analysis environmentally and biomedically significant samples