Image-inspired 3D multiphoton excited fabrication of extracellular matrix structures by modulated raster scanning

• Published in: Optics express Vol. 21; no. 21; pp. 25346 - 25355
• Main Authors: Ajeti, Visar, Lien, Chi-Hsiang, Chen, Shean-Jen, Su, Ping-Jung, Squirrell, Jayne M, Molinarolo, Katharine H, Lyons, Gary E, Eliceiri, Kevin W, Ogle, Brenda M, Campagnola, Paul J
• Format: Journal Article
• Language: English
• Published: United States 21.10.2013
• Subjects: Algorithms; Extracellular Matrix - ultrastructure; Image Enhancement - instrumentation; Image Interpretation, Computer-Assisted - instrumentation; Microscopy, Confocal - instrumentation; Microscopy, Fluorescence, Multiphoton - instrumentation
• ISSN: 1094-4087; 1094-4087
• DOI: 10.1364/oe.21.025346

Multiphoton excited photochemistry is a powerful 3D fabrication tool that produces sub-micron feature sizes. Here we exploit the freeform nature of the process to create models of the extracellular matrix (ECM) of several tissues, where the design blueprint is derived directly from high resolution optical microscopy images (e.g. fluorescence and Second Harmonic Generation). To achieve this goal, we implemented a new form of instrument control, termed modulated raster scanning, where rapid laser shuttering (10 MHz) is used to directly map the greyscale image data to the resulting protein concentration in the fabricated scaffold. Fidelity in terms of area coverage and relative concentration relative to the image data is ~95%. We compare the results to an STL approach, and find the new scheme provides significantly improved performance. We suggest the method will enable a variety of cell-matrix studies in cancer biology and also provide insight into generating scaffolds for tissue engineering.