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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 17, Iss. 8 — Apr. 13, 2009
  • pp: 6578–6583
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Optical microassembly platform for constructing reconfigurable microenvironments for biomedical studies

Peter John Rodrigo, Lóránd Kelemen, Darwin Palima, Carlo Amadeo Alonzo, Pál Ormos, and Jesper Glückstad  »View Author Affiliations


Optics Express, Vol. 17, Issue 8, pp. 6578-6583 (2009)
http://dx.doi.org/10.1364/OE.17.006578


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Abstract

Cellular development is highly influenced by the surrounding microenvironment. We propose user-reconfigurable microenvironments and bio-compatible scaffolds as an approach for understanding cellular development processes. We demonstrate a model platform for constructing versatile microenvironments by fabricating morphologically complex microstructures by two-photon polymerization (2PP) and then assembling these archetypal building blocks into various configurations using multiple, real-time configurable counterpropagating-beam (CB) traps. The demonstrated capacity for handling feature-rich microcomponents may be further developed into a generalized microassembly platform.

© 2009 Optical Society of America

1. Introduction

Earlier demonstrations have shown optically assembled 3D architectures but these were limited to using either simple microspheres [14–16

14. A. Terray, J. Oakey, and D. W. M. Marr, “Microfluidic Control Using Colloidal Devices,” Science 296, 1841–1844 (2002). [CrossRef] [PubMed]

] or generally planar, nonspherical elements [17

17. P. J. Rodrigo, L. Kelemen, C. A. Alonzo, I. R. Perch-Nielsen, J. S. Dam, P. Ormos, and J. Glückstad, “2D optical manipulation and assembly of shape-complementary planar microstructures,” Opt. Express 15, 9009–9014 (2007). [CrossRef] [PubMed]

]. Furthermore, these assembled structures required constant laser illumination to prevent constituents from drifting apart. Thus, the structural components are held in place by optical forces in the piconewton regime. In the envisioned system, feature-rich microelements are imbibed with latching mechanisms to assemble robust structures that can be maintained even upon deactivating the trapping beams to minimize parasitic effects from laser irradiation. Smaller microelements can be anchored onto larger structures to suppress Brownian dynamics. We will show that our microassembly system, based on the BioPhotonics Workstation [18

18. J. Glückstad, D. Z. Palima, J. S. Dam, and I. Perch-Nielsen, “Parallel and real-time trapping, manipulating and characterizing microscopic specimens,” Opt. Photon. News 19, 41 (2008). [CrossRef]

,19

19. H. U. Ulriksen, J. Thøgersen, S. Keiding, I. Perch-Nielsen, J. Dam, D. Z. Palima, H. Stapelfeldt, and J. Glückstad, “Independent trapping, manipulation and characterization by an all-optical biophotonics workstation,” J. Europ. Opt. Soc. Rap. Public. 3, 08034 (2008). [CrossRef]

], exhibits the necessary degrees of freedom to tackle the exacting demands arising from the increased system complexity. Experiments show that the multiple optical traps achieve sufficient degree of translational and orientational control for maneuvering the microfabricated objects in 3D.

2. Design considerations and fabrication of microscale building blocks

A microfabrication system based on two-photon photopolymerization [20

20. L. Kelemen, S. Valkai, and P. Ormos, “Integrated optical motor,” Appl. Opt. 45, 2777–2780 (2006). [CrossRef] [PubMed]

], was used to fabricate microcomponents through voxel-by-voxel solidification of an epoxy-based resin (SU8 , Michrochem, Newton, MA, USA) using tightly focused ultrashort pulses (Ti: sapphire laser, 100 fs, 80 MHz repetition rate, λ=796 nm). Setting the laser average power to 1.5 mW (focused by a 100× oil-immersion microscope objective) and using a scan speed of 8 μm/s, voxels can be solidified with minimum transverse and longitudinal dimensions of about 0.45 ± 0.1 μm and 0.8 ± 0.1 μm, respectively. In envisioned future biomedical experiments, the building blocks can be imbibed with nanometric features as needed for triggering cellular reactions and will include latching mechanisms for constructing robust microstructures while still allowing desired reconfiguration. In the present proof-of-concept demonstration, we fabricate two types of primitive microcomponents possessing complementary shape-features that enables joining them together to form hierarchical microstructures (see the scanning electron microscopy (SEM) images in Fig. 1). The microdumbbell is 13 μm long and contains identical spherical endings, 3.8 μm in diameter, connected by a cylindrical rod 2.2 μm in diameter. The dumbbells are designed to attach onto a complementary microblock (8 μm thickness and 17.5 μm side length). This is accomplished by fitting the dumbell’s spherical endings onto circular holes (diameter = 5 μm) on the sides of the microblock.

Fig. 1. Scanning electron microscope (SEM) images of 2PP-fabricated microstructures. The submicron lateral and axial resolution of resin solidification and the nanometer-precise 3D scanning paths enable 2PP to render the fine features of the microscale dumbbell (left) and its complementary primitive (right).

3. Reconfigurable microenvironment through parallel and interactive optical micromanipulation

A CW fiber laser beam (λ=1064 nm, IPG Photonics) was transformed into two matched sets of real-time user-adjustable multiple top-hat beams using spatial light modulation technology. The beam sets are coupled through facing microscope objectives (50×, NA = 0.55, Olympus) and enter from opposite sides of a sample chamber to work as counterpropagating beam traps for the fluid-borne microstructures (a small amount of surfactant prevents the particles from sticking together or to the substrate). Each counterpropagating beam (CB) trap consists of twin top-hat beams whose radii are matched to the structure to be trapped (e.g., ~4 μm radii for the dumbbell’s spherical endings). Top-hat beams evolve as they propagate and in our usual implementation of CB traps the planes where the counterpropagating beams form top-hat profiles are separated by ~40 μm. Stable three-dimensional trapping is obtained within the region where the propagated beams overlap and axial manipulation is performed by varying the power ratio between the constituent beams of a CB trap [22

22. P. J. Rodrigo, I. R. Perch-Nielsen, and J. Glückstad, “Three-dimensional forces in GPC-based counterpropagating-beam traps,” Opt. Express 14, 5812–5822 (2006). [CrossRef] [PubMed]

]. For the present experiments, the power of each constituent beam is adjustable from 0 to ~3 mW and provides the desired particle control along the beam axis.

Multiple CB traps are simultaneously configured through a computer graphical user interface and form optical potential landscapes for controlled manipulation of microcomponents. By employing a tandem of two adjustable CB traps, each dumbbell microstructure can be optically controlled and manipulated with three translational degrees of freedom (DOF) for its center-of-mass (x, y, z) and two angular DOF for its long-axis orientation (θ, ϕ), as depicted in Fig. 2(a) and demonstrated by the snapshots of video recordings from optical trapping and micromanipulation experiments (see (Figs. 2(b)–2(g)) Stable axial position control of the dumbbell primitives was achieved over a 12 μm dynamic range. Steady trapping with an extensive range of tip-tilt adjustment of the dumbbell’s long axis was done by applying varying power ratios to each lobe with programmed concurrent change in the transverse separation according to trigonometric constraints (e.g., see D and D0 in Fig. 2(e)). Stable trapping in 3D is achieved within tip-tilt ranges of θ ∈ [0, 360°] and ϕ ∈ [−50°, 50°].

Fig. 2. Stable 3D optical manipulation of a microstructure. (a) Schematic of multiple counterpropagating-beam traps for controlled manipulation of dumbbell microstructures with five degrees of freedom (x, y, z, θ, and ϕ). (b)–(d): Snapshots of controlled axial displacement of a dumbbell microstructure at fixed azimuth and zenith orientations (Media 1, 2x speed). (e)–(g): Snapshots of tip-tilt control of a dumbbell microstructure (Media 2, 2x speed). A second set of optical traps holds another dumbbell in place for reference. Overlays show relevant quantities.

Fig. 3. Snapshots from acquired video illustrating optical microassembly of reconfigurable microenvironments using 2PP-fabricated components (Media 3, 3x speed). (a) A cluster of four CB traps is selected to rotate a microblock in 2D. (b) 3D optical manipulation of dumbbell microstructures with a pair of CB traps fitting spherical lobes to holes of complementary microblock. (c) Successful assembly of all the microcomponents. (d) Grouping all CB traps to collectively manipulate the entire assembly of microscale building blocks.

4. Summary and outlook

Acknowledgments

We thank the support from the Danish Technical Scientific Research Council (FTP), the Hungarian Scientific Research Fund (grant T 046747 for P. O.) and the National Office for Research and Technology, Hungary (grant NKFP1/0007/2005 for L. K.). We thank Erzsébet Mihalik at the Department of Botany and Botanic Garden, University of Szeged, for the SEM images.

References and Links

1.

A. S. G. Curtis and M. Varde, “Control of cell behaviour: Topological factors,” J. Natl. Cancer. Inst. 33, 15–26 (1964). [PubMed]

2.

C. S. Chen, M. Mrksich, S. Huang, G. M. Whitesides, and D. E. Ingber, “Geometric control of cell life and death,” Science 276, 1425–1428 (1997). [CrossRef] [PubMed]

3.

N. Arneborg, H. Siegumfeldt, G. H. Andersen, P. Nissen, V. R. Daria, P. J. Rodrigo, and J. Glückstad, “Interactive optical trapping shows that confinement is a determinant of growth in a mixed yeast culture,” FEMS Microbiol. Lett. 245, 155–159 (2005). [CrossRef] [PubMed]

4.

J. Y. Lim and H. J. Donahue, “Cell Sensing and Response to Micro- and Nanostructured Surfaces Produced by Chemical and Topographic Patterning,” Tissue Eng. 13, 1879–1891 (2007). [CrossRef] [PubMed]

5.

M. J. Dalby, N. Gadegaard, R. Tare, A. Y. Andar, M. O. Riehle, P. Herzyk, C. D. W. Wilkinson, and R. O. C. Oreffo, “The control of human mesenchymal cell differentiation using nanoscale symmetry and disorder,” Nature Mater. 6, 997–1003 (2007). [CrossRef]

6.

B. D. MacArthur and R. O. C. Oreffo, “Bridging the gap,” Nature 433, 19 (2005). [CrossRef] [PubMed]

7.

S. Maruo, O. Nakamura, and S. Kawata, “Three-dimensional microfabrication with two-photon-absorbed photopolymerization,” Opt. Lett. 22, 132–134 (1997). [CrossRef] [PubMed]

8.

S. Kawata, H. B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices -Micromachines can be created with higher resolution using two-photon absorption,” Nature 412, 697–698 (2001). [CrossRef] [PubMed]

9.

J. F. Xing, X. -Z. Dong, W. -Q. Chen, X. -M. Duan, N. Takeyasu, T. Tanaka, and S. Kawata, “Improving spatial resolution of two-photon microfabrication by using photoinitiator with high initiating efficiency,” Appl. Phys. Lett. , 90, 131106 (2007). [CrossRef]

10.

J. L. Ifkovits and J. A. Burdick, “Review: Photopolymerizable and Degradable Biomaterials for Tissue Engineering Applications,” Tissue Eng. 13, 2369–2385 (2007). [CrossRef] [PubMed]

11.

A. Ovsianikov, S. Schlie, A. Ngezahayo, A. Haverich, and B. N. Chichkov, “Two-photon polymerization technique for microfabrication of CAD-designed 3D scaffolds from commercially available photosensitive materials,” J. Tissue Engin. Regen. Med. 1, 443–449 (2007). [CrossRef]

12.

F. Claeyssens, E. A. Hasan, A. Gaidukeviciute, D. S. Achilleos, A. Ranella, C. Reinhardt, A. Ovsianikov, X. Shizhou, C. Fotakis, M. Vamvakaki, B. N. Chichkov, and M. Farsari, “Three-Dimensional Biodegradable Structures Fabricated by Two-Photon Polymerization,” Langmuir 25, 3219–3223 (2009). [CrossRef] [PubMed]

13.

L. Li and J. T. Fourkas, “Multiphoton polymerization,” Mater. Today 10, 30–37 (2007). [CrossRef]

14.

A. Terray, J. Oakey, and D. W. M. Marr, “Microfluidic Control Using Colloidal Devices,” Science 296, 1841–1844 (2002). [CrossRef] [PubMed]

15.

Y. Roichman and D. G. Grier, “Holographic assembly of quasicrystalline photonic heterostructures,” Opt. Express 13, 5434–5439 (2005). [CrossRef] [PubMed]

16.

P. J. Rodrigo, V. R. Daria, and J. Glückstad, “Four-dimensional manipulation of colloidal particles,” Appl. Phys. Lett. 86, 074103 (2005). [CrossRef]

17.

P. J. Rodrigo, L. Kelemen, C. A. Alonzo, I. R. Perch-Nielsen, J. S. Dam, P. Ormos, and J. Glückstad, “2D optical manipulation and assembly of shape-complementary planar microstructures,” Opt. Express 15, 9009–9014 (2007). [CrossRef] [PubMed]

18.

J. Glückstad, D. Z. Palima, J. S. Dam, and I. Perch-Nielsen, “Parallel and real-time trapping, manipulating and characterizing microscopic specimens,” Opt. Photon. News 19, 41 (2008). [CrossRef]

19.

H. U. Ulriksen, J. Thøgersen, S. Keiding, I. Perch-Nielsen, J. Dam, D. Z. Palima, H. Stapelfeldt, and J. Glückstad, “Independent trapping, manipulation and characterization by an all-optical biophotonics workstation,” J. Europ. Opt. Soc. Rap. Public. 3, 08034 (2008). [CrossRef]

20.

L. Kelemen, S. Valkai, and P. Ormos, “Integrated optical motor,” Appl. Opt. 45, 2777–2780 (2006). [CrossRef] [PubMed]

21.

P Satir and S. T. Christensen, “Overview of structure and function of mammalian cilia,” Annu. Rev. Physiol. 69, 377–400 (2007). [CrossRef]

22.

P. J. Rodrigo, I. R. Perch-Nielsen, and J. Glückstad, “Three-dimensional forces in GPC-based counterpropagating-beam traps,” Opt. Express 14, 5812–5822 (2006). [CrossRef] [PubMed]

23.

I. R. Perch-Nielsen, P. J. Rodrigo, C. A. Alonzo, and J. Glückstad, “Autonomous and 3D real-time multi-beam manipulation in a microfluidic environment,” Opt. Express 14, 12199–12205 (2006). [CrossRef] [PubMed]

24.

J. Glückstad, “Sorting particles with light,” Nature Mater. 3, 9–10 (2004). [CrossRef]

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.4520) Medical optics and biotechnology : Optical confinement and manipulation
(230.4000) Optical devices : Microstructure fabrication

ToC Category:
Medical Optics and Biotechnology

History
Original Manuscript: February 18, 2009
Revised Manuscript: April 3, 2009
Manuscript Accepted: April 3, 2009
Published: April 6, 2009

Virtual Issues
Vol. 4, Iss. 6 Virtual Journal for Biomedical Optics

Citation
Peter John Rodrigo, Lóránd Kelemen, Darwin Palima, Carlo Amadeo Alonzo, Pál Ormos, and Jesper Glückstad, "Optical microassembly platform for constructing reconfigurable microenvironments for biomedical studies," Opt. Express 17, 6578-6583 (2009)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-17-8-6578


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References

  1. A. S. G. Curtis and M. Varde, "Control of cell behaviour: Topological factors," J. Natl. Cancer. Inst. 33, 15-26 (1964). [PubMed]
  2. C. S. Chen, M. Mrksich, S. Huang, G. M. Whitesides, and D. E. Ingber, "Geometric control of cell life and death," Science 276,1425-1428 (1997). [CrossRef] [PubMed]
  3. N. Arneborg, H. Siegumfeldt, G. H. Andersen, P. Nissen, V. R. Daria, P. J. Rodrigo, and J. Glückstad, "Interactive optical trapping shows that confinement is a determinant of growth in a mixed yeast culture," FEMS Microbiol. Lett. 245, 155-159 (2005). [CrossRef] [PubMed]
  4. J. Y. Lim and H. J. Donahue, "Cell Sensing and Response to Micro- and Nanostructured Surfaces Produced by Chemical and Topographic Patterning," Tissue Eng. 13, 1879-1891 (2007). [CrossRef] [PubMed]
  5. M. J. Dalby, N. Gadegaard, R. Tare, A. Y. Andar, M. O. Riehle, P. Herzyk, C. D. W. Wilkinson, and R. O. C. Oreffo, "The control of human mesenchymal cell differentiation using nanoscale symmetry and disorder," Nature Mater. 6, 997-1003 (2007). [CrossRef]
  6. B. D. MacArthur and R. O. C. Oreffo, "Bridging the gap," Nature 433, 19 (2005). [CrossRef] [PubMed]
  7. S. Maruo, O. Nakamura, and S. Kawata, "Three-dimensional microfabrication with two-photon-absorbed photopolymerization," Opt. Lett. 22, 132-134 (1997). [CrossRef] [PubMed]
  8. S. Kawata, H. B. Sun, T. Tanaka, and K. Takada, "Finer features for functional microdevices - Micromachines can be created with higher resolution using two-photon absorption," Nature 412, 697-698 (2001). [CrossRef] [PubMed]
  9. J. F. Xing, X. -Z. Dong, W. -Q. Chen, X. -M. Duan, N. Takeyasu, T. Tanaka, and S. Kawata, "Improving spatial resolution of two-photon microfabrication by using photoinitiator with high initiating efficiency," Appl. Phys. Lett.,  90, 131106 (2007). [CrossRef]
  10. J. L. Ifkovits and J. A. Burdick, "Review: Photopolymerizable and Degradable Biomaterials for Tissue Engineering Applications," Tissue Eng. 13,2369-2385 (2007). [CrossRef] [PubMed]
  11. A. Ovsianikov, S. Schlie, A. Ngezahayo, A. Haverich, and B. N. Chichkov, "Two-photon polymerization technique for microfabrication of CAD-designed 3D scaffolds from commercially available photosensitive materials," J. Tissue Engin. Regen. Med. 1, 443-449 (2007). [CrossRef]
  12. F. Claeyssens, E. A. Hasan, A. Gaidukeviciute, D. S. Achilleos, A. Ranella, C. Reinhardt, A. Ovsianikov, X. Shizhou, C. Fotakis, M. Vamvakaki, B. N. Chichkov, and M. Farsari, "Three-Dimensional Biodegradable Structures Fabricated by Two-Photon Polymerization," Langmuir 25, 3219-3223 (2009). [CrossRef] [PubMed]
  13. L. Li and J. T. Fourkas, "Multiphoton polymerization," Mater. Today 10, 30-37 (2007). [CrossRef]
  14. A. Terray, J. Oakey, and D. W. M. Marr, "Microfluidic Control Using Colloidal Devices," Science 296, 1841-1844 (2002). [CrossRef] [PubMed]
  15. Y. Roichman and D. G. Grier, "Holographic assembly of quasicrystalline photonic heterostructures," Opt. Express 13, 5434-5439 (2005). [CrossRef] [PubMed]
  16. P. J. Rodrigo, V. R. Daria, and J. Glückstad, "Four-dimensional manipulation of colloidal particles," Appl. Phys. Lett. 86, 074103 (2005). [CrossRef]
  17. P. J. Rodrigo, L. Kelemen, C. A. Alonzo, I. R. Perch-Nielsen, J. S. Dam, P. Ormos, and J. Glückstad, "2D optical manipulation and assembly of shape-complementary planar microstructures," Opt. Express 15, 9009-9014 (2007). [CrossRef] [PubMed]
  18. J. Glückstad, D. Z. Palima, J. S. Dam, and I. Perch-Nielsen, "Parallel and real-time trapping, manipulating and characterizing microscopic specimens," Opt. Photon. News 19, 41 (2008). [CrossRef]
  19. H. U. Ulriksen, J. Thøgersen, S. Keiding, I. Perch-Nielsen, J. Dam, D. Z. Palima, H. Stapelfeldt, and J. Glückstad, "Independent trapping, manipulation and characterization by an all-optical biophotonics workstation," J. Europ. Opt. Soc. Rap. Public. 3,08034 (2008). [CrossRef]
  20. L. Kelemen, S. Valkai, and P. Ormos, "Integrated optical motor," Appl. Opt. 45, 2777-2780 (2006). [CrossRef] [PubMed]
  21. P Satir, S. T. Christensen, "Overview of structure and function of mammalian cilia," Annu. Rev. Physiol. 69,377-400 (2007). [CrossRef]
  22. P. J. Rodrigo, I. R. Perch-Nielsen, and J. Glückstad, "Three-dimensional forces in GPC-based counterpropagating-beam traps," Opt. Express 14, 5812-5822 (2006). [CrossRef] [PubMed]
  23. I. R.  Perch-Nielsen, P. J.  Rodrigo, C. A.  Alonzo, and J.  Glückstad, "Autonomous and 3D real-time multi-beam manipulation in a microfluidic environment," Opt. Express 14, 12199-12205 (2006). [CrossRef] [PubMed]
  24. J. Glückstad, "Sorting particles with light," Nature Mater. 3, 9-10 (2004). [CrossRef]

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