title: Three dimensional live cell lithography.
abstract: We investigate holographic optical trapping combined with step-and-repeat maskless projection stereolithography for fine control of 3D position of living cells within a 3D microstructured hydrogel. C2C12 myoblast cells were chosen as a demonstration platform since their development into multinucleated myotubes requires linear arrangements of myoblasts. C2C12 cells are positioned in the monomer solution with multiple optical traps at 1064 nm and then encapsulated by photopolymerization of monomer via projection of a 512x512 spatial light modulator illuminated at 405 nm. High 405 nm sensitivity and complete insensitivity to 1064 nm was enabled by a lithium acylphosphinate (LAP) salt photoinitiator. These wavelengths, in addition to brightfield imaging with a white light LED, could be simultaneously focused by a single oil immersion objective. Large lateral dimensions of the patterned gel/cell structure are achieved by x and y step-and-repeat process. Large thickness is achieved through multi-layer stereolithography, allowing fabrication of precisely-arranged 3D live cell scaffolds with micron-scale structure and millimeter dimensions. Cells are shown to retain viability after the trapping and encapsulation procedure.
authors.affiliation: Department of Electrical, Computer, and Energy Engineering, University of Colorado, 1111 Engineering Drive, 422 UCB, Boulder, Colorado 80309-0422, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Linnenberger
authors.forename: Martha I
authors.initials: MI
authors.lastname: Bodine
authors.forename: Callie
authors.initials: C
authors.lastname: Fiedler
authors.forename: Justine J
authors.initials: JJ
authors.lastname: Roberts
authors.forename: Stacey C
authors.initials: SC
authors.lastname: Skaalure
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Quinn
authors.forename: Stephanie J
authors.initials: SJ
authors.lastname: Bryant
authors.forename: Michael
authors.initials: M
authors.lastname: Cole
authors.forename: Robert R
authors.initials: RR
authors.lastname: McLeod
mesh_terms: D000818:Animals; D002460:Cell Line; D004867:Equipment Design; D019544:Equipment Failure Analysis; D021621:Imaging, Three-Dimensional / Q000295:instrumentation*; D051379:Mice; D008846:Micromanipulation / Q000379:methods*; D032446:Myoblasts / Q000166:cytology* / Q000502:physiology*; D055096:Optical Devices