We present a new technique of three-dimensional tracking by way of incoherent digital holography suitable for non-scanning fluorescence microscopy. By acquiring complex holograms of a sample at different times, a difference hologram can be calculated. This result is another complex hologram representing only the information which has changed throughout the volumetric space during the time interval between one hologram and the next. We first demonstrate the advanced capability of self-interference incoherent digital holography combined with difference holography to track three-dimensional changes in a broadband, unfiltered, sunlit scene containing macroscopic continuous objects. This case is particularly remarkable due to the exceptionally short temporal coherence length and excessive build-up of noninterfering source points. We then demonstrate the ease of adaptation to microscopy, successfully converting a standard two-dimensional commercial microscope into a powerful three-dimensional tool. By overcoming these challenges, we show the feasibility and ease at which this technique may be adapted to the versatile, functional imaging of fluorescence microscopy.

Date Published: 11 March 2015
PDF: 7 pages
Proc. SPIE 9336, Quantitative Phase Imaging, 933609 (11 March 2015); doi: 10.1117/12.2078195

Published in SPIE Proceedings Vol. 9336:
Quantitative Phase Imaging
Gabriel Popescu; YongKeun Park, Editor(s)