Differential fluorescence holography

David C. Clark; Myung K. Kim

doi:10.1117/12.2078195

11 March 2015 Differential fluorescence holography

David C. Clark, Myung K. Kim

Proceedings Volume 9336, Quantitative Phase Imaging; 933609 (2015) https://doi.org/10.1117/12.2078195
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

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.

Citation Download Citation

David C. Clark and Myung K. Kim "Differential fluorescence holography", Proc. SPIE 9336, Quantitative Phase Imaging, 933609 (11 March 2015); https://doi.org/10.1117/12.2078195

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