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Microfluidic System for Transmission Electron Microscopy

Published online by Cambridge University Press:  31 August 2010

Elisabeth A. Ring  and
Niels de Jonge
Elisabeth A. Ring
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, 2215 Garland Ave., Nashville, TN 37232-0615, USA
Niels de Jonge*
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, 2215 Garland Ave., Nashville, TN 37232-0615, USA Materials Science and Technology Division, Oak Ridge National Laboratory, 1 Bethel Valley Rd., Oak Ridge, TN 37831-6064, USA
*
Corresponding author. E-mail: niels.de.jonge@vanderbilt.edu
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Abstract

We present a microfluidic system that maintains liquid flow in a specimen chamber for scanning transmission electron microscope (STEM) imaging. The specimen chamber consists of two ultrathin silicon nitride windows supported by silicon microchips. They are placed in a specimen holder that seals the sample from the vacuum in the electron microscope and incorporates tubing to and from the sample connected to a syringe pump outside the microscope. Using results obtained from fluorescence microscopy of microspheres flowing through the system, an equation to characterize the liquid flow through the system was calibrated. Gold nanoparticles of diameters of 30 and 100 nm moving in liquid were imaged with a 200 kV STEM. It was concluded that despite strong influences from Brownian motion, and sensitivity to small changes in the depth of the bypass channel, the electron microscopy flow data matched the calculated flow speed within an order of magnitude. The system allows for rapid (within a minute) liquid exchange, which can potentially be used, for example, to investigate the response of specimens, e.g., eukaryotic or bacterial cells, to certain stimuli.

Keywords

liquid flowelectron microscopynanoparticlesscanning transmission electron microscopywatermicroscopy of cells
Type
STEM Development and Applications
Information
Microscopy and Microanalysis , Volume 16 , Issue 5 , October 2010 , pp. 622 - 629
Copyright
Copyright © Microscopy Society of America 2010

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References

REFERENCES

Abrams, I.M. & McBain, J.W. (1944). A closed cell for electron microscopy. J Appl Phys 15, 607–609.CrossRefGoogle Scholar
Bao, J.B. & Harrison, D.J. (2006). Measurement of flow in microfluidic networks with micrometer-sized flow restrictors. AIChE J 52(1), 75–85.CrossRefGoogle Scholar
Crewe, A.V., Wall, J. & Langmore, J. (1970). Visibility of single atoms. Science 168, 1338–1340.CrossRefGoogle ScholarPubMed
Daulton, T.L., Little, B.J., Lowe, K. & Jones-Meehan, J. (2001). In situ environmental cell–transmission electron microscopy study of microbial reduction of chromium(VI) using electron energy loss spectroscopy. Microsc Microanal 7, 470–485.CrossRefGoogle ScholarPubMed
de Jonge, N., Peckys, D.B., Kremers, G.J. & Piston, D.W. (2009). Electron microscopy of whole cells in liquid with nanometer resolution. Proc Natl Acad Sci 106, 2159–2164.CrossRefGoogle ScholarPubMed
de Jonge, N., Peckys, D.B., Veith, G.M., Mick, S., Pennycook, S.J. & Joy, C.S. (2007). Scanning transmission electron microscopy of samples in liquid (liquid STEM). Microsc Microanal 13(S2), 242–243.CrossRefGoogle Scholar
de Jonge, N., Poirier-Demers, N., Peckys, D.B. & Drouin, D. (2010). Nanometer-resolution electron microscopy through micrometers-thick water layers. Ultramicroscopy 110(9), 1114–1119.CrossRefGoogle ScholarPubMed
Easley, C.J. (2006). Development and application of microfluidic genetic analysis systems. Dissertation in Chemistry. Charlottesville, VA: University of Virginia.Google Scholar
Einstein, A. (1905). On the motion—required by the molecular kinetic theory of heat—of small particles suspended in a stationary liquid. Ann Phys 17, 549–560.CrossRefGoogle Scholar
Kirk, S.E., Shepper, J.N. & Donald, A.M. (2009). Application of environmental scanning electron microscopy to determine biological surface structure. J Microsc 233, 205–244.CrossRefGoogle ScholarPubMed
Liu, K.L., Wu, C.C., Huang, Y.J., Peng, H.L., Chang, H.Y., Chang, P., Hsu, L. & Yew, T.R. (2008). Novel microchip for in situ TEM imaging of living organisms and bio-reactions in aqueous conditions. Lab Chip 8, 1915–1921.CrossRefGoogle ScholarPubMed
Longwell, P.A. (1966). Mechanics of Fluid Flow. New York: McGraw Hill.Google Scholar
Nishiyama, H., Sugo, M., Ogura, T., Maruyama, Y., Koizumi, M., Mio, K., Kitamura, S. & Sato, C. (2010). Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film. J Struct Biol 169(3), 438–439.CrossRefGoogle ScholarPubMed
Parsons, D.F., Matricardi, V.R., Moretz, R.C. & Turner, J.N. (1974). Electron microscopy and diffraction of wet unstained and unfixed biological objects. Adv Biol Med Phys 15, 161–270.CrossRefGoogle ScholarPubMed
Peckys, D.B., Veith, G.M., Joy, D.C. & de Jonge, N. (2009). Nanoscale imaging of whole cells using a liquid enclosure and a scanning transmission electron microscope. PLoS One 4(12), e8214.CrossRefGoogle Scholar
Thiberge, S., Nechushtan, A., Sprinzak, D., Gileadi, O., Behar, V., Zik, O., Chowers, Y., Michaeli, S., Schlessinger, J. & Moses, E. (2004). Scanning electron microscopy of cells and tissues under fully hydrated conditions. Proc Natl Acad Sci 101(10), 3346.CrossRefGoogle ScholarPubMed
Williamson, M.J., Tromp, R.M., Vereecken, P.M., Hull, R. & Ross, F.M. (2003). Dynamic microscopy of nanoscale cluster growth at the solid-liquid interface. Nat Mater 2, 532–536.CrossRefGoogle ScholarPubMed
Xiao, Y., Patolsky, F., Katz, E., Hainfeld, J.F. & Willner, I. (2003). “Plugging into enzymes”: Nanowiring of redox enzymes by a gold nanoparticle. Science 299, 1877–1881.CrossRefGoogle ScholarPubMed
Zheng, H., Smith, R.K., Jun, Y.W., Kisielowski, C., Dahmen, U. & Alivisatos, A.P. (2009). Observation of single colloidal platinum nanocrystal growth trajectories. Science 324(5932), 1309–1312.CrossRefGoogle ScholarPubMed
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