Bubble Jet agent release cartridge for chemical single cell stimulation
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We present a new method for the distinct specific chemical stimulation of single cells and small cell clusters within their natural environment. By single-drop release of chemical agents with droplets in size of typical cell diameters (d <30 μm) on-demand micro gradients can be generated for the specific manipulation of single cells. A single channel and a double channel agent release cartridge with integrated fluidic structures and integrated agent reservoirs are shown, tested, and compared in this publication. The single channel setup features a fluidic structure fabricated by anisotropic etching of silicon. To allow for simultaneous release of different agents even though maintaining the same device size, the second type comprises a double channel fluidic structure, fabricated by photolithographic patterning of TMMF. Dispensed droplet volumes are V = 15 pl and V = 10 pl for the silicon and the TMMF based setups, respectively. Utilizing the agent release cartridges, the application in biological assays was demonstrated by hormone-stimulated premature bud formation in Physcomitrella patens and the individual staining of one single L 929 cell within a confluent grown cell culture.
KeywordsSingle cell stimulation Drop-on-demand InkJet Agent release Pyscomitrella patens L-929
We gratefully thank the German Research Foundation (DFG, ZE 527/4 and EXC 294) for financial support of this project. We also appreciate the good cooperation with the Cleanroom Service Center at IMTEK.
- Cellectricon, Dynaflow®Pro II System – High Quality Automated Patch Clamping (Cellectricon AB, Mölndal, Sweden). http://www.cellectricon.se. Accessed Feb. 2011
- H.G. Drexler, et al. DSMZ Catalogue of Human and Animal Cell Lines, Eighth Edition. (2001)Google Scholar
- R. Elmqvist, Measuring instrument of the recording type, US Patent 2566443, 1951Google Scholar
- Eppendorf, Microinjectors FemtoJet and FemtoJet express Product Sheet (Eppendorf AG Hamburg, Germany). http://www.eppendorf.de. Accessed Feb. 2011
- P. Jonas, Fast application of agonists to isolated membrane patches. In Single-Channel Recording, eds. B. Sakmann & E. Neher (Plenum press, 1995), pp. 231–243Google Scholar
- H.P. Le, J. Imag. Sci. Tech. 42, 49–62 (1998)Google Scholar
- Pam3Cys-SKKKK(Rhodamin), (EMC microcollections GmbH, Tübingen, Germany). http://www.microcollections.de. Accessed Feb. 2011
- R. Reski, Botanica Acta 111, 1–15 (1998)Google Scholar
- U. Stöhr, P. Vulto, P. Hoppe, G. Urban, H. Reinecke, Journal of Micro/Nanolithography, MEMS, and MOEMS. 7, 033009 (2008)Google Scholar
- N. Wangler et al., Proceedings of Thirteenth International Conference on Miniaturized Systems for Chemistry and Life Sciences November 1–5. (2009), 679–81Google Scholar
- N. Wangler et al., J. Micromech. Microeng. 21(095009), 9 (2011)Google Scholar