3D imaging is demanding technology required in fluorescence microscopy. Even though holography is a powerful technique, it could not be used easily in fluorescence microscopy because of low coherence of fluorescence light. Lately, several incoherent holographic methods such as scanning holography, Fresnel in coherent correlation holography (FINCH), and self-interference incoherent digital holography (SIDH) have been proposed. However, these methods have many problems to be overcome for practical applications. For example, DC term removal, twin image ambiguity, and phase unwrapping problems need to be resolved. Off-axis holography is a straightforward solution which can solve most of these problems. We built an off-axis SIDH system for fluorescence imaging, and investigated various conditions and requirements for practical holographic fluorescence microscopy. Our system is based on a modified Michelson interferometer with a flat mirror at one arm and a curved mirror at the other arm of the interferometer. We made a phantom 3D fluorescence object made of 2 single-mode fibers coupled to a single red LED source to mimic 2 fluorescence point sources distributed by a few tens of micrometers apart. A cooled EM-CCD was used to take holograms of these fiber ends which emit only around 180 nW power.
|Title of host publication||Practical Holography XXXI|
|Subtitle of host publication||Materials and Applications|
|Editors||Hans I. Bjelkhagen, V. Michael Bove|
|Publication status||Published - 2017|
|Event||SPIE Conference on Practical Holography XXXI: Materials and Applications - San Francisco, United States|
Duration: 2017 Jan 30 → 2017 Feb 1
|Name||Proceedings of SPIE - The International Society for Optical Engineering|
|Other||SPIE Conference on Practical Holography XXXI: Materials and Applications|
|Period||17/1/30 → 17/2/1|
Bibliographical noteFunding Information:
This work was financially supported by the MEST through the National Research Foundation of Korea (Grant No. 2012R1A4A1029061), the Center for Advanced Meta-Materials (CAMM) funded by the Ministry of Science, ICT and Future Planning as Global Frontier Project (CAMM-2014M3A6B3063712), the Technology Innovation Program (10062417) funded by the Ministry of Trade, industry and Energy (MI), and the Ministry of Education Science and Technology of Korea through the BK21 program.
© 2017 SPIE.
All Science Journal Classification (ASJC) codes
- Electronic, Optical and Magnetic Materials
- Condensed Matter Physics
- Computer Science Applications
- Applied Mathematics
- Electrical and Electronic Engineering