Special Section on Coherent Raman Imaging Techniques and Biomedical Applications

Coherent anti-Stokes Raman scattering microscopy of human smooth muscle cells in bioengineered tissue scaffolds

[+] Author Affiliations
Christian Brackmann, Annika Enejder

Chalmers University of Technology, Molecular Microscopy, Department of Chemical and Biological Engineering, SE-412 96 Göteborg, Sweden

Maricris Esguerra, Daniel Olausson, Dick Delbro

Sahlgrenska Academy at University of Gothenburg, Institute of Clinical Sciences, Department of Surgery, SE-413 45 Göteborg, Sweden

Alexandra Krettek

Sahlgrenska Academy at University of Gothenburg, Institute of Medicine, Department of Internal Medicine, SE-405 30 Göteborg, Sweden

Paul Gatenholm

Chalmers University of Technology, Polymer Science, Department of Chemical and Biological Engineering, SE-412 96 Göteborg, Sweden

J. Biomed. Opt. 16(2), 021115 (February 04, 2011). doi:10.1117/1.3534782
History: Received June 13, 2010; Revised December 03, 2010; Accepted December 15, 2010; Published February 04, 2011; Online February 04, 2011
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The integration of living, human smooth muscle cells in biosynthesized cellulose scaffolds was monitored by nonlinear microscopy toward contractile artificial blood vessels. Combined coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) microscopy was applied for studies of the cell interaction with the biopolymer network. CARS microscopy probing CH2-groups at 2845 cm−1 permitted three-dimensional imaging of the cells with high contrast for lipid-rich intracellular structures. SHG microscopy visualized the fibers of the cellulose scaffold, together with a small signal obtained from the cytoplasmic myosin of the muscle cells. From the overlay images we conclude a close interaction between cells and cellulose fibers. We followed the cell migration into the three-dimensional structure, illustrating that while the cells submerge into the scaffold they extrude filopodia on top of the surface. A comparison between compact and porous scaffolds reveals a migration depth of <10 μm for the former, whereas the porous type shows cells further submerged into the cellulose. Thus, the scaffold architecture determines the degree of cell integration. We conclude that the unique ability of nonlinear microscopy to visualize the three-dimensional composition of living, soft matter makes it an ideal instrument within tissue engineering.

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© 2011 Society of Photo-Optical Instrumentation Engineers (SPIE)

Citation

Christian Brackmann ; Maricris Esguerra ; Daniel Olausson ; Dick Delbro ; Alexandra Krettek, et al.
"Coherent anti-Stokes Raman scattering microscopy of human smooth muscle cells in bioengineered tissue scaffolds", J. Biomed. Opt. 16(2), 021115 (February 04, 2011). ; http://dx.doi.org/10.1117/1.3534782


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