Abstract

Holographic microscopes are emerging as suitable tools for in situ diagnostics and environmental monitoring, providing high-throughput, label-free, quantitative imaging capabilities through small and compact devices. In-line holographic microscopes can be realized at contained costs, trading off complexity in the phase retrieval process and being limited to sparse samples. Here we present a 3D printed, cost effective and field portable off-axis holographic microscope based on the concept of holographic microfluidic slide. Our scheme removes complexity from the reconstruction process, as phase retrieval is non iterative and obtainable by hologram demodulation. The configuration we introduce ensures flexibility in the definition of the optical scheme, exploitable to realize modular devices with different features. We discuss trade-offs and design rules of thumb to follow for developing DH microscopes based on the proposed solution. Using our prototype, we image flowing marine microalgae, polystyrene beads, E.coli bacteria and microplastics. We detail the effect on the performance and costs of each parameter, design, and hardware choice, guiding readers toward the realization of optimized devices that can be employed out of the lab by non-expert users for point of care testing.

© 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

Full Article  |  PDF Article
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References

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2019 (4)

2018 (10)

Y. Rivenson, Y. Zhang, H. Günaydın, D. Teng, and A. Ozcan, “Phase recovery and holographic image reconstruction using deep learning in neural networks,” Light: Sci. Appl. 7(2), 17141 (2018).
[Crossref]

N. Patel, V. Trivedi, S. Mahajan, V. Chhaniwal, C. Fournier, S. Lee, B. Javidi, and A. Anand, “Wavefront Division Digital Holographic Microscopy,” Biomed. Opt. Express 9(6), 2779–2784 (2018).
[Crossref]

V. Bianco, P. Memmolo, M. Leo, S. Montresor, C. Distante, M. Paturzo, P. Picart, B. Javidi, and P. Ferraro, “Strategies for reducing speckle noise in digital holography,” Light: Sci. Appl. 7(1), 48 (2018).
[Crossref]

C. H. Clausen, M. Dimaki, C. V. Bertelsen, G. E. Skands, R. Rodriguez-Trujillo, J. Dahl Thomsen, and W. E. Svendsen, “Bacteria Detection and Differentiation Using Impedance Flow Cytometry,” Sensors 18(10), 3496 (2018).
[Crossref]

P. Paiè, R. M. Vázquez, R. Osellame, F. Bragheri, and A. Bassi, “Microfluidic Based Optical Microscopes on Chip,” Cytometry, Part A 93(10), 987–996 (2018).
[Crossref]

M. Mugnano, P. Memmolo, L. Miccio, F. Merola, V. Bianco, A. Bramanti, A. Gambale, R. Russo, I. Andolfo, A. Iolascon, and P. Ferraro, “Label-Free Optical Marker for Red-Blood-Cell Phenotyping of Inherited Anemias,” Anal. Chem. 90(12), 7495–7501 (2018).
[Crossref]

Z. Göröcs, M. Tamamitsu, V. Bianco, P. Wolf, S. Roy, K. Shindo, K. Yanny, Y. Wu, H. C. Koydemir, Y. Rivenson, and A. Ozcan, “A deep learning-enabled portable imaging flow cytometer for cost-effective, high-throughput, and label-free analysis of natural water samples,” Light: Sci. Appl. 7(1), 66 (2018).
[Crossref]

M. Villone, P. Memmolo, F. Merola, M. Mugnano, L. Miccio, P. Maffettone, and P. Ferraro, “Full-angle tomographic phase microscopy of flowing quasi-spherical cells,” Lab Chip 18(1), 126–131 (2018).
[Crossref]

Y. Zhang, H. Koydemir, M. Shimogawa, S. Yalcin, A. Guziak, T. Liu, I. Oguz, Y. Huang, B. Bai, Y. Luo, Y. Luo, Z. Wei, H. Wang, V. Bianco, B. Zhang, R. Nadkarni, K. Hill, and A. Ozcan, “Motility-based label-free detection of parasites in bodily fluids using holographic speckle analysis and deep learning,” Light: Sci. Appl. 7(1), 108 (2018).
[Crossref]

F. Merola, P. Memmolo, V. Bianco, M. Paturzo, M. G. Mazzocchi, and P. Ferraro, “Searching and identifying microplastics in marine environment by digital holography,” Eur. Phys. J. Plus 133(9), 350 (2018).
[Crossref]

2017 (5)

L. A. Philips, D. B. Ruffner, F. C. Cheong, J. M. Blusewicz, P. Kasimbeg, B. Waisi, J. R. McCutcheon, and D. G. Grier, “Holographic characterization of contaminants in water: Differentiation of suspended particles in heterogeneous dispersions,” Water Res. 122, 431–439 (2017).
[Crossref]

V. Bianco, B. Mandracchia, V. Marchesano, V. Pagliarulo, F. Olivieri, S. Coppola, M. Paturzo, and P. Ferraro, “Endowing a plain fluidic chip with micro-optics: a holographic microscope slide,” Light Sci. Appl. 6(9), e17055 (2017).
[Crossref]

S. Vashist, “Point-of-Care Diagnostics: Recent Advances and Trends,” Biosensors 7(4), 62 (2017).
[Crossref]

M. Daloglu and A. Ozcan, “Computational imaging of sperm locomotion,” Biol. Reprod. 97(2), 182–188 (2017).
[Crossref]

B. Mandracchia, V. Bianco, Z. Wang, M. Mugnano, A. Bramanti, M. Paturzo, and P. Ferraro, “Holographic microscope slide in a spatio-temporal imaging modality for reliable 3D cell counting,” Lab Chip 17(16), 2831–2838 (2017).
[Crossref]

2016 (1)

V. Vespini, S. Coppola, M. Todino, M. Paturzo, V. Bianco, S. Grilli, and P. Ferraro, “Forward electrohydrodynamic inkjet printing of optical microlenses on microfluidic devices,” Lab Chip 16(2), 326–333 (2016).
[Crossref]

2015 (4)

X. Li, X. Zhu, Q. Zhou, H. Wang, and K. Ni, “Low-cost lithography for fabrication of one-dimensional diffraction gratings by using laser diodes,” Proc. SPIE 9624, 962408 (2015).
[Crossref]

M. Rahlves, M. Rezem, K. Boroz, S. Schlangen, E. Reithmeier, and B. Roth, “Flexible, fast, and low-cost production process for polymer based diffractive optics,” Opt. Express 23(3), 3614 (2015).
[Crossref]

Y. Jo, J. Jung, M. Kim, H. Park, S. Kang, and Y. Park, “Label-free identification of individual bacteria using Fourier transform light scattering,” Opt. Express 23(12), 15792–15805 (2015).
[Crossref]

F. Merola, P. Memmolo, L. Miccio, V. Bianco, M. Paturzo, and P. Ferraro, “Diagnostic Tools for Lab-on-Chip Applications Based on Coherent Imaging Microscopy,” Proc. IEEE 103(2), 192–204 (2015).
[Crossref]

2014 (4)

E. Sanchez-Ortiga, A. Doblas, G. Saavedra, M. Martínez-Corral, and J. Garcia-Sucerquia, “Off-axis digital holographic microscopy: practical design parameters for operating at diffraction limit,” Appl. Opt. 53(10), 2058–2066 (2014).
[Crossref]

P. Y. Liu, L. K. Chin, W. Ser, T. C. Ayi, P. H. Yap, T. Bourouina, and Y. Leprince-Wang, “An optofluidic imaging system to measure the biophysical signature of single waterborne bacteria,” Lab Chip 14(21), 4237–4243 (2014).
[Crossref]

N. C. Pégard, M. L. Toth, M. Driscoll, and J. W. Fleischer, “Flow-scanning optical tomography,” Lab Chip 14(23), 4447–4450 (2014).
[Crossref]

P. Memmolo, L. Miccio, F. Merola, O. Gennari, P. A. Netti, and P. Ferraro, “3D morphometry of red blood cells by digital holography,” Cytometry, Part A 85(12), 1030–1036 (2014).
[Crossref]

2013 (6)

C. M. Rochman and M. A. Browne, “Classify plastic waste as hazardous,” Nature 494(7436), 169–171 (2013).
[Crossref]

D. Hoornweg, P. Bhada-Tata, and C. Kennedy, “Environment: Waste production must peak this century,” Nature 502(7473), 615–617 (2013).
[Crossref]

R. Guo, B. Yao, P. Gao, J. Min, M. Zhou, J. Han, X. Yu, X. Yu, M. Lei, S. Yan, Y. Yang, D. Dan, and T. Ye, “Off-axis digital holographic microscopy with LED illumination based on polarization filtering,” Appl. Opt. 52(34), 8233–8238 (2013).
[Crossref]

J. C. Petruccelli, L. Tian, and G. Barbastathis, “The transport of intensity equation for optical path length recovery using partially coherent illumination,” Opt. Express 21(12), 14430–14441 (2013).
[Crossref]

J. M. Swiecicki, O. Sliusarenko, and D. B. Weibel, “From swimming to swarming: Escherichia coli cell motility in two-dimensions,” Integr. Biol. 5(12), 1490–1494 (2013).
[Crossref]

Y. Cotte, F. Toy, P. Jourdain, N. Pavillon, D. Boss, P. Magistretti, P. Marquet, and C. Depeursinge, “Marker-free phase nanoscopy,” Nat. Photonics 7(2), 113–117 (2013).
[Crossref]

2012 (4)

2011 (1)

2010 (1)

O. Mudanyali, D. Tseng, C. Oh, S. O. Isikman, I. Sencan, W. Bishara, C. Oztoprak, S. Seo, B. Khademhosseini, and A. Ozcan, “Compact, light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications,” Lab Chip 10(11), 1417–1428 (2010).
[Crossref]

2009 (1)

E. V. Armbrust, “The life of diatoms in the world’s oceans,” Nature 459(7244), 185–192 (2009).
[Crossref]

2006 (1)

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigation of living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11(3), 034005 (2006).
[Crossref]

2001 (1)

W. Xu, M. H. Jericho, I. A. Meinertzhagen, and H. J. Kreuzer, “Digital in-line holography for biological applications,” Proc. Natl. Acad. Sci. U. S. A. 98(20), 11301–11305 (2001).
[Crossref]

Anand, A.

Andolfo, I.

M. Mugnano, P. Memmolo, L. Miccio, F. Merola, V. Bianco, A. Bramanti, A. Gambale, R. Russo, I. Andolfo, A. Iolascon, and P. Ferraro, “Label-Free Optical Marker for Red-Blood-Cell Phenotyping of Inherited Anemias,” Anal. Chem. 90(12), 7495–7501 (2018).
[Crossref]

Armbrust, E. V.

E. V. Armbrust, “The life of diatoms in the world’s oceans,” Nature 459(7244), 185–192 (2009).
[Crossref]

Ayi, T. C.

P. Y. Liu, L. K. Chin, W. Ser, T. C. Ayi, P. H. Yap, T. Bourouina, and Y. Leprince-Wang, “An optofluidic imaging system to measure the biophysical signature of single waterborne bacteria,” Lab Chip 14(21), 4237–4243 (2014).
[Crossref]

Bai, B.

Y. Zhang, H. Koydemir, M. Shimogawa, S. Yalcin, A. Guziak, T. Liu, I. Oguz, Y. Huang, B. Bai, Y. Luo, Y. Luo, Z. Wei, H. Wang, V. Bianco, B. Zhang, R. Nadkarni, K. Hill, and A. Ozcan, “Motility-based label-free detection of parasites in bodily fluids using holographic speckle analysis and deep learning,” Light: Sci. Appl. 7(1), 108 (2018).
[Crossref]

Barbastathis, G.

Bassi, A.

P. Paiè, R. M. Vázquez, R. Osellame, F. Bragheri, and A. Bassi, “Microfluidic Based Optical Microscopes on Chip,” Cytometry, Part A 93(10), 987–996 (2018).
[Crossref]

Bertelsen, C. V.

C. H. Clausen, M. Dimaki, C. V. Bertelsen, G. E. Skands, R. Rodriguez-Trujillo, J. Dahl Thomsen, and W. E. Svendsen, “Bacteria Detection and Differentiation Using Impedance Flow Cytometry,” Sensors 18(10), 3496 (2018).
[Crossref]

Bhada-Tata, P.

D. Hoornweg, P. Bhada-Tata, and C. Kennedy, “Environment: Waste production must peak this century,” Nature 502(7473), 615–617 (2013).
[Crossref]

Bian, Z.

Bianco, V.

V. Bianco, P. Memmolo, M. Leo, S. Montresor, C. Distante, M. Paturzo, P. Picart, B. Javidi, and P. Ferraro, “Strategies for reducing speckle noise in digital holography,” Light: Sci. Appl. 7(1), 48 (2018).
[Crossref]

M. Mugnano, P. Memmolo, L. Miccio, F. Merola, V. Bianco, A. Bramanti, A. Gambale, R. Russo, I. Andolfo, A. Iolascon, and P. Ferraro, “Label-Free Optical Marker for Red-Blood-Cell Phenotyping of Inherited Anemias,” Anal. Chem. 90(12), 7495–7501 (2018).
[Crossref]

Z. Göröcs, M. Tamamitsu, V. Bianco, P. Wolf, S. Roy, K. Shindo, K. Yanny, Y. Wu, H. C. Koydemir, Y. Rivenson, and A. Ozcan, “A deep learning-enabled portable imaging flow cytometer for cost-effective, high-throughput, and label-free analysis of natural water samples,” Light: Sci. Appl. 7(1), 66 (2018).
[Crossref]

Y. Zhang, H. Koydemir, M. Shimogawa, S. Yalcin, A. Guziak, T. Liu, I. Oguz, Y. Huang, B. Bai, Y. Luo, Y. Luo, Z. Wei, H. Wang, V. Bianco, B. Zhang, R. Nadkarni, K. Hill, and A. Ozcan, “Motility-based label-free detection of parasites in bodily fluids using holographic speckle analysis and deep learning,” Light: Sci. Appl. 7(1), 108 (2018).
[Crossref]

F. Merola, P. Memmolo, V. Bianco, M. Paturzo, M. G. Mazzocchi, and P. Ferraro, “Searching and identifying microplastics in marine environment by digital holography,” Eur. Phys. J. Plus 133(9), 350 (2018).
[Crossref]

B. Mandracchia, V. Bianco, Z. Wang, M. Mugnano, A. Bramanti, M. Paturzo, and P. Ferraro, “Holographic microscope slide in a spatio-temporal imaging modality for reliable 3D cell counting,” Lab Chip 17(16), 2831–2838 (2017).
[Crossref]

V. Bianco, B. Mandracchia, V. Marchesano, V. Pagliarulo, F. Olivieri, S. Coppola, M. Paturzo, and P. Ferraro, “Endowing a plain fluidic chip with micro-optics: a holographic microscope slide,” Light Sci. Appl. 6(9), e17055 (2017).
[Crossref]

V. Vespini, S. Coppola, M. Todino, M. Paturzo, V. Bianco, S. Grilli, and P. Ferraro, “Forward electrohydrodynamic inkjet printing of optical microlenses on microfluidic devices,” Lab Chip 16(2), 326–333 (2016).
[Crossref]

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F. Merola, P. Memmolo, L. Miccio, V. Bianco, M. Paturzo, and P. Ferraro, “Diagnostic Tools for Lab-on-Chip Applications Based on Coherent Imaging Microscopy,” Proc. IEEE 103(2), 192–204 (2015).
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P. Memmolo, L. Miccio, F. Merola, O. Gennari, P. A. Netti, and P. Ferraro, “3D morphometry of red blood cells by digital holography,” Cytometry, Part A 85(12), 1030–1036 (2014).
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https://www.lynceetec.com/

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Supplementary Material (1)

NameDescription
» Visualization 1       Media 1 is a hologram sequence showing a diatom rotating while flowing along the microfluidic channel, recorded largely out of focus.

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Figures (9)

Fig. 1.
Fig. 1. Compact HSM (a) 3D model and (b) corresponding 3D printed prototype, realized as per configuration in panel d. (c) Schematic representations of the setup “Configuration A” and (d) “Configuration B”. R: reference beam; O: object beam; wg: grating width; s: separation between grating and microchannel; dGL: distance grating-lens; dLC: distance lens-camera.
Fig. 2.
Fig. 2. Phase contrast mapping using the proposed configuration A. (a,b) Marine algae (diatoms) fixed on a glass slide. (a) Pseudo 3D view of the diatoms inside the red rectangles in (b) after DH reconstruction. $\Delta x$ and $\Delta y$ denote the pixel size in the image plane. (b) Bright-field 5x microscope image of the slide containing the set of imaged diatoms. (c) Thickness of E-coli bacteria inside a microfluidic channel is measured. White circles indicate the positions where bacteria are automatically detected.
Fig. 3.
Fig. 3. DH imaging of diatoms fixed on a glass slide using the proposed Configuration B. (a) Digital hologram shows the test sample largely out of focus. (b) Amplitude reconstruction of the samples after DH numerical refocusing. (c) Bright-field microscope image (5x magnification) of the glass slide.
Fig. 4.
Fig. 4. (Visualization 1) The HSM device shown in Fig. 1 was used to image objects of interest for environmental monitoring. (a-b) Diatoms flowing inside a microfluidic channel. (a) Out of focus DHs of a flowing and rotating diatom are superposed on the same image to show the movement. The corresponding phase-contrast maps after refocusing each DH are shown in the insets. (b) 3D rendering of the rotating diatom in (a), obtained as a result of the SFS algorithm. (c) Amplitude reconstructions of a flowing diatom in different time instants show the movement of the alga due to the flow. The corresponding phase contrast maps are shown in the insets. The bright-field microscope image of the same diatom class is shown for the sake of comparison in the top-right corner of (c). (d-g) Imaging microplastics using the proposed field portable DH microscope of Fig. 1. (d) T(z) vs. z. (e). Out of focus DH of a PVC plastic item. (f) Amplitude DH reconstruction at z maximizing T(z). (g) Bright-field microscope image of PVC plastics.
Fig. 5.
Fig. 5. Recording geometry for the setup Configuration A, in lensless modality.
Fig. 6.
Fig. 6. Changes in the recording geometry for the setup Configuration A by adding a lens.
Fig. 7.
Fig. 7. Selection of distance dOC in setup Configuration A.
Fig. 8.
Fig. 8. Refocusing holograms of polystyrene micro-beads captured using configuration A. (a) Digital hologram. (b) Bright-field microscope image of the same beads. (c) Tamura coefficient, $T(z )$ vs. z. DH amplitude reconstructions at different z are shown in the insets.
Fig. 9.
Fig. 9. Details of the PMMA chip structure. Dimensions are expressed in mm.

Tables (2)

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Table 1. Retail price in Euro for the main hardware components of the HSM. The total cost of the 3D printed microscope in Fig. 1 is estimated to be lower than 800€ for the production of one single device, and lower than 300€ for large production volumes.

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Table 2. Relation between the main optical and geometrical parameters in Configuration B. The combination selected in our work is highlighted in blue.

Equations (9)

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h = | O ( x , y ) | 2 + | R ( x , y ) | 2 + O ( x , y ) | R ( x , y ) | e j k 0 s i n θ x + O ( x , y ) | R ( x , y ) | e j k 0 s i n θ x
T ( z ) = σ [ | P { H + 1 ; z } | ] μ [ | P { H + 1 ; z } | ]
{ α tan α = ( N Δ p 2 + w O 2 ) 1 d O C , β = a sin ( λ g )
α + β < θ N y q β < λ 2 Δ p ( N Δ p 2 + w O 2 ) 1 d O C
α = ( N Δ p 2 + w 2 ) 1 d
{ d O C tan β w g 0 d O C ( w o 2 + s + w g 2 ) cot β
M f n Δ p sin ( β ) λ
d L C min = f ( M f + 1 )
M o b j = f f p o b j ,

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