Microscopy and Microanalysis

Techniques, Software, and Instrumentation Development

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

Yingjun Yana1a2, Liwei Jianga1, Karl J. Aufderheidea3, Gus A. Wrighta1, Alexander Terekhova4, Lino Costaa4, Kevin Qina1a2, W. Tyler McCleerya5, John J. Fellensteina6, Alessandro Ustionea7, J. Brian Robertsona1, Carl Hirschie Johnsona1, David W. Pistona7, M. Shane Hutsona5a8, John P. Wikswoa5a7a8a9, William Hofmeistera4a8 and Chris Janetopoulosa1a2a8 c1

a1 Department of Biological Sciences, Vanderbilt University, Nashville, TN 37232, USA

a2 Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN 37232, USA

a3 Department of Biology, Texas A&M University, College Station, TX 77843, USA

a4 Center for Laser Applications, University of Tennessee Space Institute, Tullahoma, TN 37388, USA

a5 Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 37232, USA

a6 Vanderbilt Machine Shop, Vanderbilt University, Nashville, TN 37232, USA

a7 Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA

a8 Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN 37232, USA

a9 Department of Biomedical Engineering, Vanderbilt University, Nashville, TN 37232, USA

Abstract

A microcompressor is a precision mechanical device that flattens and immobilizes living cells and small organisms for optical microscopy, allowing enhanced visualization of sub-cellular structures and organelles. We have developed an easily fabricated device, which can be equipped with microfluidics, permitting the addition of media or chemicals during observation. This device can be used on both upright and inverted microscopes. The apparatus permits micrometer precision flattening for nondestructive immobilization of specimens as small as a bacterium, while also accommodating larger specimens, such as Caenorhabditis elegans, for long-term observations. The compressor mount is removable and allows easy specimen addition and recovery for later observation. Several customized specimen beds can be incorporated into the base. To demonstrate the capabilities of the device, we have imaged numerous cellular events in several protozoan species, in yeast cells, and in Drosophila melanogaster embryos. We have been able to document previously unreported events, and also perform photobleaching experiments, in conjugating Tetrahymena thermophila.

(Received September 18 2013)

(Accepted November 06 2013)

Key words

  • microcompression;
  • microfluidics;
  • laser machining;
  • pyramidal wells;
  • Paramecium tetraurelia ;
  • Saccharomyces cerevisiae ;
  • polydimethylsiloxane

Correspondence

c1 Corresponding author. E-mail: c.janetopoulos@vanderbilt.edu

Footnotes

  Both authors contributed equally to this study.