Detection, imaging, and kinetics of sub-micron organelles of chondrocytes by multiple beam interference microscopy

Narahari V. Joshi; Honorio Medina; J.M. Barboza; Gladys Colantuoni; Maritza Quintero

doi:10.1117/12.527544

13 July 2004 Detection, imaging, and kinetics of sub-micron organelles of chondrocytes by multiple beam interference microscopy

Narahari V. Joshi, Honorio Medina, J.M. Barboza, Gladys Colantuoni, Maritza Quintero

Author Affiliations +

Proceedings Volume 5324, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XI; (2004) https://doi.org/10.1117/12.527544
Event: Biomedical Optics 2004, 2004, San Jose, CA, United States

Abstract

Chondrocytes, obtained from testosterone treated human articular cartilage, were examined by a recently developed Multiple Beam Interference Microscopy (MBIM) attached to a confocal set up, Video-enhanced differential interference microphotography and also by cinematography. In the MBIM, the intensity of the transmitted pattern is given by the Airy function which increases the contrast dramatically as the coefficient of the reflectance of the parallel plates increases. Moreover, in this configuration, the beam passes several times through a specific organelle and increases its optical path difference both because of the increase in the trajectory and refractive index (high density) of the organelle. The improved contrast enhances the resolving power of the system and makes visible several structural details of sub micron dimensions like nucleolus, retraction fibers, podia, etc. which are not possible to reveal with such a clarity by conventional techniques such as bright field, phase contrast or DIC. This technique permits to detect the oscillatory and rotational motions of unstained cilia for the first time. The frequency of oscillations was found to be 0.8 Hz.

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Narahari V. Joshi, Honorio Medina, J.M. Barboza, Gladys Colantuoni, and Maritza Quintero "Detection, imaging, and kinetics of sub-micron organelles of chondrocytes by multiple beam interference microscopy", Proc. SPIE 5324, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XI, (13 July 2004); https://doi.org/10.1117/12.527544

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KEYWORDS

Microscopy

Video

Optical fibers

Luminescence

Confocal microscopy

Spectral resolution

CCD cameras

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