Multiwavelength Digital Holographic Imaging and Phase Unwrapping of Protozoa Using Custom Fiji Plug-ins
Multiwavelength digital holographic microscopy (DHM) has been used to improve phase reconstructions of digital holograms by reducing 2π phase ambiguities. However, most samples used as test images have been solid or adhered to a surface, making it easy to determine focal planes and correct for chrom...
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| Published in: | Frontiers in physics Vol. 7 |
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Frontiers Media S.A
05-07-2019
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| Abstract | Multiwavelength digital holographic microscopy (DHM) has been used to improve phase reconstructions of digital holograms by reducing 2π phase ambiguities. However, most samples used as test images have been solid or adhered to a surface, making it easy to determine focal planes and correct for chromatic aberration. In this study we apply 3-wavelength off-axis DHM to swimming protozoa containing distinct spectral features such as chlorophyll and carotenoids. We reconstruct the holograms into amplitude and phase images using the angular spectrum method. Methods for noise subtraction, chromatic aberration correction, and image registration are presented for both amplitude and phase. Approaches to phase unwrapping are evaluated and compared to expected results from simulated holograms. The algorithms used are implemented in plug-ins using the open source Fiji platform and are available for use, significantly expanding the open-source software available for DHM. |
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| AbstractList | Multiwavelength digital holographic microscopy (DHM) has been used to improve phase reconstructions of digital holograms by reducing 2π phase ambiguities. However, most samples used as test images have been solid or adhered to a surface, making it easy to determine focal planes and correct for chromatic aberration. In this study we apply 3-wavelength off-axis DHM to swimming protozoa containing distinct spectral features such as chlorophyll and carotenoids. We reconstruct the holograms into amplitude and phase images using the angular spectrum method. Methods for noise subtraction, chromatic aberration correction, and image registration are presented for both amplitude and phase. Approaches to phase unwrapping are evaluated and compared to expected results from simulated holograms. The algorithms used are implemented in plug-ins using the open source Fiji platform and are available for use, significantly expanding the open-source software available for DHM. |
| Author | Hanczarek, Iulia Wallace, J. Kent Nadeau, Jay Cohoe, David |
| Author_xml | – sequence: 1 givenname: David surname: Cohoe fullname: Cohoe, David – sequence: 2 givenname: Iulia surname: Hanczarek fullname: Hanczarek, Iulia – sequence: 3 givenname: J. Kent surname: Wallace fullname: Wallace, J. Kent – sequence: 4 givenname: Jay surname: Nadeau fullname: Nadeau, Jay |
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| Cites_doi | 10.1007/978-1-4419-7793-9 10.1109/AERO.2019.8742091 10.1088/0957-0233/13/9/201 10.1364/AO.45.000851 10.1016/j.jsb.2012.03.008 10.1364/OE.23.017367 10.1117/1.JBO.18.3.036007 10.1016/j.pocean.2018.09.002 10.1371/journal.pone.0147700 10.1364/OL.30.000468 10.3390/app8030444 10.1117/1.JBO.19.4.046009 10.1364/AO.45.000451 10.1364/OE.15.007231 10.1371/journal.pone.0051041 10.1038/nrmicro3567 10.1371/journal.pone.0030912 10.1007/BF00425189 10.1371/journal.pone.0045044 10.1364/OL.28.001141 10.1364/OL.30.002706 10.1364/AO.54.006410 10.3791/56343 10.1364/OL.43.001943 10.1364/OL.24.000291 10.1364/oe.14.004300 10.1016/j.ijleo.2017.04.091 10.1021/j100858a002 10.1038/s41566-018-0253-x 10.1038/s41598-018-25922-7 10.1364/OL.31.000178 |
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| References | Khmaladze (B7) 2012; 178 Turko (B28) 2018; 43 Jourdain (B6) 2012; 7 Isikman (B34) 2012; 7 Saalfeld (B29) 2008 Son (B3) 2015; 13 Colomb (B25) 2014; 39 Wallace (B23) 2019 Park (B14) 2018; 12 Singhal (B31) 1968; 72 Kuhn (B17) 2007; 15 Parshall (B18) 2006; 45 Zahalsky (B21) 1962; 42 Pavillon (B8) 2012; 7 Marquet (B1) 2005; 30 Kim (B10) 2011 Lehmuskero (B32) 2018; 168 Nehmetallah (B11) 2016; 25 De Nicola (B16) 2005; 30 Charriere (B33) 2006; 31 Cuche (B13) 1999; 24 Wallace (B22) 2015; 23 Boss (B2) 2013; 18 Goldstein (B27) 2013 Matsuda (B19) 2018; 8 Piedrahita-Quintero (B30) 2017; 140 Lindensmith (B4) 2016; 11 Bedrossian (B5) 2017; 129 Schnars (B9) 2002; 13 Foltinova (B20) 1994; 49 Kus (B35) 2014; 19 Piedrahita-Quintero (B24) 2015; 54 Colomb (B26) 2006; 45 Liu (B12) 2018; 8 Gass (B15) 2003; 28 |
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| Title | Multiwavelength Digital Holographic Imaging and Phase Unwrapping of Protozoa Using Custom Fiji Plug-ins |
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