Microscope enabling multimodality imaging, angle-resolved scattering, and scattering spectroscopy.
We present the design, construction, and initial characterization of a multifunctional imaging/scattering spectroscopy system built around a commercial inverted microscope platform. The system enables co-registered brightfield, Fourier-filtered darkfield, and fluorescence imaging; monochromatic angle-resolved scattering measurements; and white-light wavelength-resolved scattering spectroscopy from the same field of view. A fiber-based illumination system provides illumination-wavelength flexibility and a good approximation to a point source. The performance of the system in its various data acquisition modes is experimentally verified using fluorescent microspheres. This multifunctional instrument provides a platform for studies on adherent cells from which the biophysical implications of subcellular light scattering can be studied in conjunction with sensitive fluorescence-based techniques.
Cottrell
W J
WJ
Institute of Optics and Departments of Physics and of Imaging Sciences, University of Rochester, Rochester, NY 14642, USA.
Wilson
J D
JD
Foster
T H
TH
D008027:Light; D008856:Microscopy, Fluorescence / Q000379:methods; D015388:Organelles / Q000737:chemistry*; D012542:Scattering, Radiation; D013057:Spectrum Analysis / Q000379:methods