Microscopy with spatial filtering for sorting particles and monitoring subcellular morphology

Jing-Yi Zheng; Zhen Qian; Robert M. Pasternack; Nada N. Boustany

doi:10.1117/12.809555

12 February 2009 Microscopy with spatial filtering for sorting particles and monitoring subcellular morphology

Jing-Yi Zheng, Zhen Qian, Robert M. Pasternack, Nada N. Boustany

Proceedings Volume 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII; 71821G (2009) https://doi.org/10.1117/12.809555
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Optical scatter imaging (OSI) was developed to non-invasively track real-time changes in particle morphology with submicron sensitivity in situ without exogenous labeling, cell fixing, or organelle isolation. For spherical particles, the intensity ratio of wide-to-narrow angle scatter (OSIR, Optical Scatter Image Ratio) was shown to decrease monotonically with diameter and agree with Mie theory. In living cells, we recently reported this technique is able to detect mitochondrial morphological alterations, which were mediated by the Bcl-xL transmembrane domain, and could not be observed by fluorescence or differential interference contrast images. Here we further extend the ability of morphology assessment by adopting a digital micromirror device (DMD) for Fourier filtering. When placed in the Fourier plane the DMD can be used to select scattering intensities at desired combination of scattering angles. We designed an optical filter bank consisting of Gabor-like filters with various scales and rotations based on Gabor filters, which have been widely used for localization of spatial and frequency information in digital images and texture analysis. Using a model system consisting of mixtures of polystyrene spheres and bacteria, we show how this system can be used to sort particles on a microscopic slide based on their size, orientation and aspect ratio. We are currently applying this technique to characterize the morphology of subcellular organelles to help understand fundamental biological processes.

Citation Download Citation

Jing-Yi Zheng, Zhen Qian, Robert M. Pasternack, and Nada N. Boustany "Microscopy with spatial filtering for sorting particles and monitoring subcellular morphology", Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 71821G (12 February 2009); https://doi.org/10.1117/12.809555

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KEYWORDS

Particles

Optical spheres

Digital micromirror devices

Optical filters

Image filtering

Mirrors

Spatial frequencies

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