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Collinear optical coherence and confocal fluorescence microscopies for tissue engineering

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