Abstract
A novel contact printing method utilizing a sacrificial layer of polyacrylic acid (PAA) was developed to selectively modify the upper surfaces of arrayed microstructures. The method was characterized by printing polystyrene onto SU-8 microstructures to create an improved substrate for a cell-based microarray platform. Experiments measuring cell growth on SU-8 arrays modified with polystyrene and fibronectin demonstrated improved growth of NIH 3T3 (93% vs. 38%), HeLa (97% vs. 77%), and HT1080 (76% vs. 20%) cells relative to that for the previously used coating method. In addition, use of the PAA sacrificial layer permitted the printing of functionalized polystyrene, carboxylate polystyrene nanospheres, and silica nanospheres onto the arrays in a facile manner. Finally, a high concentration of extracellular matrix materials (ECM), such as collagen (5 mg/mL) and gelatin (0.1%), was contact-printed onto the array structures using as little as 5 μL of the ECM reagent and without the formation of a continuous film bridge across the microstructures. Murine embryonic stem cells cultured on arrays printed with this gelatin hydrogel remained in an undifferentiated state indicating an adequate surface gelatin layer to maintain these cells over time.
Similar content being viewed by others
References
Khademhosseini A, Langer R, Borenstein J, Vacanti JP (2006) Microscale technologies for tissue engineering and biology. PNAS 103:2480–2487
Paguirigan AL, Beebe DJ (2008) Microfluidics meet cell biology: bridging the gap by validation and application of microscale techniques for cell biological assays. BioEssays 30:811–821
Gomez F (2008) Biological applications of microfluidics. Wiley, Hoboken
Yarmush ML, King KR (2009) Living-cell microarrays. Annu Rev Biomed Eng 11:235–257
Sims CE, Allbritton NL (2007) Analysis of single mammalian cells on-chip. Lab Chip 7:423–440
Fernandes TG, Diogo MM, Clark DS, Dordick JS, Cabral JMS (2009) High-throughput cellular microarray platforms: applications in drug discovery, toxicology and stem cell research. Trends Biotechnol 27:342–349
Pirone DM, Chen CS (2004) Strategies for engineering the adhesive microenvironment. J Mammary Gland Biol Neoplasia 9:405–417
Rehfeldt F, Engler AJ, Eckhardt A, Ahmed F, Discher DE (2007) Cell responses to the mechanochemical microenvironment—implications for regenerative medicine and drug delivery. Adv Drug Deliv Rev 59:1329–1339
Wang Y, Bachman M, Sims CE, Li GP, Allbritton NL (2006) Simple photografting method to chemically modify and micropattern the surface of SU-8 photoresist. Langmuir 22:2719–2725
Dittami GM, Ayliffe HE, King CS, Rabbitt RD (2008) A multilayer MEMS platform for single-cell electric impedance spectroscopy and electrochemical analysis. J Microelectromech Syst 17:850–862
Wakamoto Y, Inoue I, Moriguchi H, Yasuda K (2001) Analysis of single-cell differences by use of an on-chip microculture system and optical trapping. Fresen J Anal Chem 371:276–281
Wu M-H, Cai H, Xu X, Urban JPG, Cui Z-F, Cui Z (2005) A SU-8/PDMS hybrid microfluidic device with integrated optical fibers for online monitoring. Biomed Microdevices 7:323–329
Umehara S, Wakamoto Y, Inoue I, Yasuda K (2003) On-chip single-cell microcultivation assay for monitoring environmental effects on isolated cells. Biochem Biophys Res Commun 305:534–540
Chronis N, Lee LP (2004) Electrothermally activated SU-8 microgripper for single cell manipulation in solution. J Microelectromech Syst 14:857–863
Calleja M, Tamayo J, Nordström M, Boisen A (2006) Low-noise polymeric nanomechanical biosensors. Appl Phys Lett 88:113901–113903
Johansson A, Calleja M, Rasmussen PA, Boisen A (2005) SU-8 cantilever sensor system with integrated readout. Sens Actuators A Phys 123–124:111–115
Shew BY, Kuo CH, Huang YC, Tsai YH (2005) UV-LIGA interferometer biosensor based on the SU-8 optical waveguide. Sens Actuators A Phys 120:383–389
Wang L, Wu Z-Z, Xu B, Zhao Y, Kisaalita WS (2009) SU-8 microstructure for quasi-three-dimensional cell-based biosensing. Sens Actuators B Chem 140:349–355
Wu ZZ, Zhao Y, Kisaalita WS (2006) Interfacing SH-SY5Y human neuroblastoma cells with SU-8 microstructures. Colloids Surf B Biointerfaces 52:14–21
Wang Y, Young G, Bachman M, Sims CE, Li GP, Allbritton NL (2007) Collection and expansion of single cells and colonies released from a micropallet array. Anal Chem 79:2359–2366
Kotzar G, Freas M, Abel P, Fleischman A, Roy S, Zorman C, Moran JM, Melzak J (2002) Evaluation of MEMS materials of construction for implantable medical devices. Biomater 23:2737–2750
Voskerician G, Shive MS, Shawgo RS, von Recum H, Anderson JM, Cima MJ, Langer R (2003) Biocompatibility and biofouling of MEMS drug delivery devices. Biomater 24:1959–1967
Grayson ACR, Shawgo RS, Johnson AM, Flynn NT, Li Y, Cima MJ, Langer RA (2004) BioMEMS review: MEMS technology for physiologically integrated devices. Proc IEEE 92:6–21
Weisenberg BA, Mooradian DL (2002) Hemocompatibility of materials used in microelectromechanical systems: platelet adhesion and morphology in vitro. J Biomed Mater Res 60:283–291
Stangegaard M, Wang Z, Kutter JP, Dufva M, Wolff A (2006) Whole genome expression profiling using DNA microarray for determining biocompatibility of polymeric surfaces. Mol Biosyst 2:421–428
Hennemeyer M, Walther F, Kerstan S, Schürzinger K, Gigler AM, Stark RW (2008) Cell proliferation assays on plasma activated SU-8. Microelectron Engr 85:1298–1301
Tao SL, Popat KC, Norman JJ, Desai TA (2008) Surface modification of SU-8 for enhanced biofunctionality and nonfouling properties. Langmuir 24:2631–2636
Vernekar VN, Cullen DK, Fogleman N, Choi Y, Garcia AJ, Allen MG, Brewer GJ, LaPlaca MC (2009) SU-8 2000 rendered cytocompatible for neuronal bioMEMS applications. J Biomed Mater Res A 89:138–151
Pai JH, Wang Y, Salazar GT, Sims CE, Bachman M, Li GP, Allbritton NL (2007) Photoresist with low fluorescence for bioanalytical applications. Anal Chem 79:8774–8780
Wang Y, Sims CE, Marc P, Bachman M, Li GP, Allbritton NL (2006) Micropatterning of living cells on a heterogeneously wetted surface. Langmuir 22:8257–8262
Hu S, Ren X, Bachman M, Sims CE, Li GP, Allbritton NL (2004) Tailoring the surface properties of poly(dimethylsiloxane) microfluidic devices. Langmuir 20:5569–5574
Stevens MP (1999) Polymer chemistry: an introduction. Oxford University Press, London
Salazar GT, Wang Y, Young G, Bachman M, Sims CE, Li GP, Allbritton NL (2007) Micropallet arrays for the separation of single, adherent cells. Anal Chem 79:682–687
Shadpour H, Sims CE, Thresher RJ, Allbritton NL (2009) Sorting and expansion of murine embryonic stem cell colonies using micropallet arrays. Cytom A 75:121–129
Quinto-Su PA, Salazar GT, Sims CE, Allbritton NL, Venugopalan V (2008) Mechanism of pulsed laser microbeam release of SU-8 2100 polymer micropallets for the collection and separation of adherent cells. Anal Chem 80:4675–4679
Linder V, Gates BD, Ryan D, Parviz BA, Whitesides GM (2005) Water-soluble sacrificial layers for surface micromachining. Small 1:730–736
Ramsey WS, Hertl W, Nowlan ED, Binkowski NJ (1984) Surface treatments and cell attachment. In Vitr 20:802–808
Yamamoto A, Mishima S, Maruyama N, Sumita M (2000) Quantitative evaluation of cell attachment to glass, polystyrene, and fibronectin- or collagen-coated polystyrene by measurement of cell adhesive shear force and cell detachment energy. J Biomed Mater Res 50:114–124
Steele JG, Dalton BA, Johnson G, Underwood PA (1995) Adsorption of fibronectin and vitronectin onto Primaria and tissue culture polystyrene and relationship to the mechanism of initial attachment of human vein endothelial cells and BHK-21 fibroblasts. Biomater 16:1057–1067
Heo J, Lee JS, Chu IS, Takahama Y, Thorgeirsson SS (2005) Spontaneous differentiation of mouse embryonic stem cells in vitro: characterization by global gene expression profiles. Biochem Biophys Res Commun 332:1061–1069
Acknowledgments
This research was supported by NIH (EB007612 and HG004843). The authors thank Chapel Hill Analytical and Nanofabrication Laboratory (CHANL) for providing access to the facility’s instrumentation. The authors also thank Dr. Yuli Wang for valuable discussions and Colleen Phillips and Jonathan Clark for technical support.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary materials
Below is the link to the electronic supplementary material.
ESM 1
(PDF 1178 kb)
Rights and permissions
About this article
Cite this article
Xu, W., Luikart, A.M., Sims, C.E. et al. Contact printing of arrayed microstructures. Anal Bioanal Chem 397, 3377–3385 (2010). https://doi.org/10.1007/s00216-010-3728-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-010-3728-2