Imaging of small particles using wide-field confocal microscopy

Stephen P. Morgan; N. B. E. Sawyer; Michael G. Somekh; Chung Wah See; B. Y. Shekunov; E. Astrakharchik

doi:10.1117/12.518774

13 October 2003 Imaging of small particles using wide-field confocal microscopy

Stephen P. Morgan, N. B. E. Sawyer, Michael G. Somekh, Chung Wah See, B. Y. Shekunov, E. Astrakharchik

Proceedings Volume 5068, Saratov Fall Meeting 2002: Optical Technologies in Biophysics and Medicine IV; (2003) https://doi.org/10.1117/12.518774
Event: Saratov Fall Meeting 2002 Laser Physics and Photonics, Spectroscopy, and Molecular Modeling III; Coherent Optics of Ordered and Random Media III, 2002, Saratov, Russian Federation

Abstract

Particle measurement is important in many applications such as the manufacture of drugs and paints, and aerosols. In bioimaging there is interest understanding the imaging of nanoparticles and subcellular scatterers. We present in this paper a wide field, phase measuring confocal microscope that can be used for such measurements. The wide field confocal response is obtained by illuminating both sample and reference arms of an interferometric microscope with nominally identical speckle patterns. When the speckle patterns are highly correlated the interference is significant. Contributions from out of focus planes result in uncorrelated speckle patterns and no interference. This provides a wide field confocal response. High speed measurements are enabled by parallel phase stepping using polarization optics. We have also developed a vector diffraction microscope model, using Mie theory as a scattering function, to validate the images of small particles. Correctly scaling the amplitudes of the unscattered and scattered electric fields enables co-polar transmission imaging to be modeled. Finally it is demonstrated that the phase is a more sensitive measurement of particle size than the amplitude.

Citation Download Citation

Stephen P. Morgan, N. B. E. Sawyer, Michael G. Somekh, Chung Wah See, B. Y. Shekunov, and E. Astrakharchik "Imaging of small particles using wide-field confocal microscopy", Proc. SPIE 5068, Saratov Fall Meeting 2002: Optical Technologies in Biophysics and Medicine IV, (13 October 2003); https://doi.org/10.1117/12.518774

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