Abstract

Tissue engineered medical products (TEMPs) are often three-dimensional (3D) hybrid materials consisting of a porous scaffold upon which the tissue is grown. However, monitoring of the developing tissue deep within the scaffold is hampered by its turbidity. We have sought new ways to probe the interior of the scaffold with the same resolution as conventional laser scanning confocal microscopy but with greater sensitivity. We present a novel application of optical coherence microscopy (OCM) by combining it with confocal fluorescence microscopy (CFM) to gather simultaneous structural and functional information on TEMPs in a registered fashion. In this work, we describe the collinear OCM and CFM instrument. We demonstrate the utility of this dual-mode technique for TEMPs by imaging fluorescently stained osteoblasts cultured in a polymeric TEMP.

©2003 Optical Society of America

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References

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  1. J. M. Anderson, L. G. Cima, and S. G. Eskinet al., “Tissue engineering in cardiovascular disease - a report,” J. Biomed. Mater. Res. 29, 1473–1476 (1995).
    [Crossref]
  2. J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
    [Crossref]
  3. S. Chinn and E. Swanson, “Blindness limitations in optical coherence domain reflectometry,” Electron. Lett. 29, 2025–2027 (1993).
    [Crossref]
  4. F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).
  5. M. Attawia, J. Devin, and C. Laurencin, “Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,” J. Biomed. Mat. Res. 29, 843–848 (1995).
    [Crossref]
  6. M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
    [Crossref] [PubMed]
  7. F. Guzman and J. K. Barton, “Dual OCT/spectroscopy system for identification of vascular pathology,” Proc. SPIE Int. Soc. Opt. Eng. 4244, 413–414 (2001).
  8. E. Beaurepaire, L. Moreaux, F. Amblard, and J. Mertz, “Combined scanning optical coherence and two-photon-excited fluorescence microscopy,” Opt. Lett. 24, 969–971 (2000).
    [Crossref]
  9. S. Demos, M. Staggs, K. Minoshima, and J. Fujimoto, “Characterization of laser induced damage sites in optical components,” Opt. Express 10, 1444–1450 (2002). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-25-1444
    [Crossref] [PubMed]
  10. Identification of a commercial product is made only to facilitate experimental reproducibility and to adequately describe experimental procedure. In no case does it imply endorsement by NIST or imply that it is necessarily the best product for the experimental procedure.
  11. All uncertainties expressed in accordance to: B. Taylor and C. Kuyatt, NIST Technical Note 1297, 1994 and are based on one standard deviation.
  12. G. Tearney, M. Brezinski, J. Southern, B. Bouma, M. Hee, and J. Fujimoto, “Determination of the refractive-index of highly scattering human tissue by optical coherence tomography,” Opt. Lett. 20, 2258–2260 (1995).
    [Crossref] [PubMed]
  13. J. O. Iroh, “Poly(ε-caprolactone)” in Polymer Data Handbook, J. E. Mark, ed. (Oxford University Press, New York. 1999).
  14. N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
    [Crossref]
  15. http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable2.html

2003 (1)

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

2002 (2)

S. Demos, M. Staggs, K. Minoshima, and J. Fujimoto, “Characterization of laser induced damage sites in optical components,” Opt. Express 10, 1444–1450 (2002). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-25-1444
[Crossref] [PubMed]

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

2001 (1)

F. Guzman and J. K. Barton, “Dual OCT/spectroscopy system for identification of vascular pathology,” Proc. SPIE Int. Soc. Opt. Eng. 4244, 413–414 (2001).

2000 (2)

E. Beaurepaire, L. Moreaux, F. Amblard, and J. Mertz, “Combined scanning optical coherence and two-photon-excited fluorescence microscopy,” Opt. Lett. 24, 969–971 (2000).
[Crossref]

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

1996 (1)

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

1995 (3)

M. Attawia, J. Devin, and C. Laurencin, “Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,” J. Biomed. Mat. Res. 29, 843–848 (1995).
[Crossref]

G. Tearney, M. Brezinski, J. Southern, B. Bouma, M. Hee, and J. Fujimoto, “Determination of the refractive-index of highly scattering human tissue by optical coherence tomography,” Opt. Lett. 20, 2258–2260 (1995).
[Crossref] [PubMed]

J. M. Anderson, L. G. Cima, and S. G. Eskinet al., “Tissue engineering in cardiovascular disease - a report,” J. Biomed. Mater. Res. 29, 1473–1476 (1995).
[Crossref]

1993 (1)

S. Chinn and E. Swanson, “Blindness limitations in optical coherence domain reflectometry,” Electron. Lett. 29, 2025–2027 (1993).
[Crossref]

Afting, M.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Amblard, F.

Amis, E.

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

Anderson, J. M.

J. M. Anderson, L. G. Cima, and S. G. Eskinet al., “Tissue engineering in cardiovascular disease - a report,” J. Biomed. Mater. Res. 29, 1473–1476 (1995).
[Crossref]

Attawia, M.

M. Attawia, J. Devin, and C. Laurencin, “Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,” J. Biomed. Mat. Res. 29, 843–848 (1995).
[Crossref]

Barton, J. K.

F. Guzman and J. K. Barton, “Dual OCT/spectroscopy system for identification of vascular pathology,” Proc. SPIE Int. Soc. Opt. Eng. 4244, 413–414 (2001).

Beaurepaire, E.

Bjerkvig, R.

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

Bouma, B.

Brezinski, M.

Chinn, S.

S. Chinn and E. Swanson, “Blindness limitations in optical coherence domain reflectometry,” Electron. Lett. 29, 2025–2027 (1993).
[Crossref]

Cima, L. G.

J. M. Anderson, L. G. Cima, and S. G. Eskinet al., “Tissue engineering in cardiovascular disease - a report,” J. Biomed. Mater. Res. 29, 1473–1476 (1995).
[Crossref]

Demos, S.

Devin, J.

M. Attawia, J. Devin, and C. Laurencin, “Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,” J. Biomed. Mat. Res. 29, 843–848 (1995).
[Crossref]

Elgendy, H.

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

Eskin, S. G.

J. M. Anderson, L. G. Cima, and S. G. Eskinet al., “Tissue engineering in cardiovascular disease - a report,” J. Biomed. Mater. Res. 29, 1473–1476 (1995).
[Crossref]

Fujimoto, J.

Guzman, F.

F. Guzman and J. K. Barton, “Dual OCT/spectroscopy system for identification of vascular pathology,” Proc. SPIE Int. Soc. Opt. Eng. 4244, 413–414 (2001).

Hee, M.

Iroh, J. O.

J. O. Iroh, “Poly(ε-caprolactone)” in Polymer Data Handbook, J. E. Mark, ed. (Oxford University Press, New York. 1999).

Izatt, J.

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

Karim, A.

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

Kobayashi, K.

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

Kulkarni, M.

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

Kuyatt, C.

All uncertainties expressed in accordance to: B. Taylor and C. Kuyatt, NIST Technical Note 1297, 1994 and are based on one standard deviation.

Laurencin, C.

M. Attawia, J. Devin, and C. Laurencin, “Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,” J. Biomed. Mat. Res. 29, 843–848 (1995).
[Crossref]

Lund-Johansen, M.

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

Mertz, J.

Minoshima, K.

Moreaux, L.

Nasseri, B.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Pomerantseva, I.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Read, T.-A.

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

Seed, B.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Simon, C.

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

Sivak, M.

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

Southern, J.

Staggs, M.

Stock, U.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Swanson, E.

S. Chinn and E. Swanson, “Blindness limitations in optical coherence domain reflectometry,” Electron. Lett. 29, 2025–2027 (1993).
[Crossref]

Taylor, B.

All uncertainties expressed in accordance to: B. Taylor and C. Kuyatt, NIST Technical Note 1297, 1994 and are based on one standard deviation.

Tearney, G.

Thorsen, F.

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

Tona, A.

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

Tysnes, B. B.

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

Vacanti, J.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Wang, H.-S.

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

Washburn, N.

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

Cell Transplantation. (1)

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, “Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,” Cell Transplantation. 9, 773–783 (2000).

Electron. Lett. (1)

S. Chinn and E. Swanson, “Blindness limitations in optical coherence domain reflectometry,” Electron. Lett. 29, 2025–2027 (1993).
[Crossref]

IEEE J. Sel. Top. Quantum Electron. (1)

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, “Optical coherence tomography and microscopy in gastrointestinal tissues,” IEEE J. Sel. Top. Quantum Electron. 2, 1017–1028 (1996).
[Crossref]

J. Biomed. Mat. Res. (2)

M. Attawia, J. Devin, and C. Laurencin, “Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,” J. Biomed. Mat. Res. 29, 843–848 (1995).
[Crossref]

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, “Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,” J. Biomed. Mat. Res. 60, 20–29 (2002).
[Crossref]

J. Biomed. Mater. Res. (1)

J. M. Anderson, L. G. Cima, and S. G. Eskinet al., “Tissue engineering in cardiovascular disease - a report,” J. Biomed. Mater. Res. 29, 1473–1476 (1995).
[Crossref]

Opt. Express (1)

Opt. Lett. (2)

Proc. SPIE Int. Soc. Opt. Eng. (1)

F. Guzman and J. K. Barton, “Dual OCT/spectroscopy system for identification of vascular pathology,” Proc. SPIE Int. Soc. Opt. Eng. 4244, 413–414 (2001).

Tissue Eng. (1)

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, “Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,” Tissue Eng. 9, 137–141 (2003).
[Crossref] [PubMed]

Other (4)

Identification of a commercial product is made only to facilitate experimental reproducibility and to adequately describe experimental procedure. In no case does it imply endorsement by NIST or imply that it is necessarily the best product for the experimental procedure.

All uncertainties expressed in accordance to: B. Taylor and C. Kuyatt, NIST Technical Note 1297, 1994 and are based on one standard deviation.

J. O. Iroh, “Poly(ε-caprolactone)” in Polymer Data Handbook, J. E. Mark, ed. (Oxford University Press, New York. 1999).

http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable2.html

Supplementary Material (1)

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Figures (4)

Fig. 1.
Fig. 1. Experimental apparatus for collinear OCM/ CFM. BP: band pass filter; CL: collimating lens; CM: cold mirror; Demod: Demodulator; SOAS: semiconductor optical amplifier source; DBS: dichroic beam splitter; LP: Long pass filter; NDF: Neutral density filter; NF: Notch filter; PM: Polarization maintaining 50/50 coupler; Pol: In-line polarizer; PS: Polarization splitter; PZTFM: Piezoelectic fiber

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Fig. 2.
Fig. 2. OCM (A.) and CFM (B.) images 145 µm from surface of PCL cultured for 10 weeks with fetal chick osteoblasts.

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Fig. 3.
Fig. 3. Scanning electron micrograph of PCL scaffold. PCL was co-extruded with poly-ethyleneoxide (PEO) at 0.50 mass fraction. The scaffold was immersed in solvent to dissolve the PEO, and the remaining PCL was subject to a 30 min anneal at 75 °C.

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Fig. 4.
Fig. 4. (0.72 MB) Movie of merged and registered OCM and CFM images of the cultured PCL scaffold.

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