Integrated Raman- and angular-scattering microscopy.

A microscopy system has been constructed that is capable of simultaneously acquiring both traditional Raman spectra as well as angle-resolved elastic-scattering patterns using a single focused laser spot less than 10 mum wide. The elastic-scattering signal was analyzed by generalized Lorenz-Mie theory, representing what we believe to be the first experimental validation of the theory's prediction of angular backscatter from single spheres. The microscope system exhibits 3 nm precision in predicting sphere diameters, while simultaneously yielding high-quality Raman signals. Applications to single cell analysis are envisioned.

[1]  Nitish V Thakor,et al.  Calcium-induced alterations in mitochondrial morphology quantified in situ with optical scatter imaging. , 2002, Biophysical journal.

[2]  Chenyang Xu,et al.  Wavelength-dependent scattering in spectroscopic optical coherence tomography. , 2005, Optics express.

[3]  Changhuei Yang,et al.  Determination of particle size by using the angular distribution of backscattered light as measured with low-coherence interferometry. , 2002, Journal of the Optical Society of America. A, Optics, image science, and vision.

[4]  J P Freyer,et al.  Polarized angular dependent spectroscopy of epithelial cells and epithelial cell nuclei to determine the size scale of scattering structures. , 2002, Journal of biomedical optics.

[5]  T. McConnell,et al.  Activation-dependent phases of T cells distinguished by use of optical tweezers and near infrared Raman spectroscopy. , 2005, Journal of immunological methods.

[6]  G Gouesbet,et al.  Scattering of laser beams by Mie scatter centers: numerical results using a localized approximation. , 1986, Applied optics.

[7]  Irving Itzkan,et al.  Confocal light absorption and scattering spectroscopic microscopy. , 2007, Applied optics.

[8]  T. Foster,et al.  Microscope enabling multimodality imaging, angle-resolved scattering, and scattering spectroscopy. , 2007, Optics letters.

[9]  A. Wax,et al.  Rapid, depth-resolved light scattering measurements using Fourier domain, angle-resolved low coherence interferometry. , 2004, Optics express.

[10]  J. Lock,et al.  Improved Gaussian beam-scattering algorithm. , 1995, Applied optics.

[11]  Thomas H Foster,et al.  Light scattering from intact cells reports oxidative-stress-induced mitochondrial swelling. , 2005, Biophysical journal.

[12]  J. Koenig,et al.  Laser Raman scattering in uniaxially oriented atactic polystyrene , 1978 .