August 2012: This Month in JoVE

Summary

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and mirrors to refract and direct light to optical sensors. The August 2012 issue of JoVE (Journal of Visualized Experiments) is marked by the third publication from the Ozcan Lab (University of California, Los Angeles) on their lens-free "on-chip" microscopy platform, which they have pioneered.

Protocol

Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children

Peiling Yap^{1, 2}, Thomas Fürst^{1, 2}, Ivan Müller^{1, 2}, Susi Kriemler^{1, 2}, Jürg Utzinger^{1, 2}, Peter Steinmann^{1, 2
1}Department of Epidemiology and Public Health, Swiss Tropical and Public Health Institute, Basel, Switzerland, ²University of Basel, Basel, Switzerland

Chronic infection with soil-transmitted helminths (STHs) causes malabsorption, stunting, and wasting in the growing child. Hence, it is plausible that these infections also reduce the physical fitness of children. Here, we visualize two techniques for the diagnosis of STHs and the 20-meter shuttle run test for assessing children's physical fitness.

In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development

Ben Yang^1,*, Lauren B. Geary^1,*, Yong-Chao Ma^{1, 2
1}Northwestern University Feinberg School of Medicine, Children's Hospital of Chicago Research Center, ²Departments of Pediatrics, Neurology and Physiology, Northwestern University Feinberg School of Medicine
^* These authors contributed equally

To assess the function and the regulation of genes during the development of midbrain dopaminergic neurons, we describe a method that involves in ovo electroporation of plasmid DNA constructs into embryonic chick ventral midbrain dopaminergic neuron progenitors. This technique can be used to achieve efficient expression of genes of interest to study different aspects of midbrain development and dopaminergic neuron differentiation.

Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions

Anutosh Ganguly, Hong Zhang, Ritu Sharma, Sean Parsons, Kamala D. Patel
Department of Physiology and Pharmacology, University of Calgary

This article first describes a procedure for isolating human endothelial cells from umbilical veins and then shows how to use these cells to examine neutrophil transmigration under flow conditions. By using a low-volume flow chamber made from a polymer with the optical characteristics of glass, live-cell fluorescent imaging of rare cell populations is also possible.

Construction and Testing of Coin Cells of Lithium Ion Batteries

Archana Kayyar¹, Jiajia Huang¹, Mojtaba Samiee¹, Jian Luo^{1, 2
1}School of Materials Science and Engineering, Clemson University, ²Center for Optical Materials Science and Engineering Technologies, Clemson University

A protocol to construct and test coin cells of lithium ion batteries is described. The specific procedures of making a working electrode, preparing a counter electrode, assembling a cell inside a glovebox and testing the cell are presented.

Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts

Karen H. Martin, Karen E. Hayes, Elyse L. Walk, Amanda Gatesman Ammer, Steven M. Markwell, Scott A. Weed
Department of Neurobiology and Anatomy, Program in Cancer Cell Biology, Mary Babb Randolph Cancer Center, West Virginia University

We describe the prototypical method for producing microscope coverslips coated with fluorescent gelatin for visualizing invadopodia-mediated matrix degradation. Computational techniques using available software are presented for quantifying the resultant levels of matrix proteolysis by single cells within a mixed population and for multicellular groups encompassing entire microscopic fields.

Lensfree On-chip Tomographic Microscopy employing Multi-Angle Illumination and Pixel Super-Resolution

Serhan O. Isikman¹, Waheb Bishara¹, Aydogan Ozcan^{1, 2, 3
1}Electrical Engineering Department, University of California, Los Angeles , ²Bioengineering Department, University of California, Los Angeles , ³California NanoSystems Institute, University of California, Los Angeles

Lensfree optical tomography is a three-dimensional microscopy technique that offers a spatial resolution of <1 μm × <1 μm × <3 μm in x, y and z dimensions, respectively, over a large imaging-volume of 15-100 mm³, which can be particularly useful for integration with lab-on-a-chip platforms.

An Analytical Tool that Quantifies Cellular Morphology Changes from Three-dimensional Fluorescence Images

Carolina L. Haass-Koffler^{1, 2}, Mohammad Naeemuddin¹, Selena E. Bartlett^{1, 3
1}Medications Development, Ernest Gallo Clinic and Research Center, University of California, San Francisco , ²Clinical Pharmacology and Experimental Therapeutics, University of California, San Francisco , ³Translational Research Institute and the Institute for Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia

We developed a software platform that utilizes Imaris Neuroscience, ImarisXT and MATLAB to measure the changes in morphology of an undefined shape taken from three-dimensional confocal fluorescence of single cells. This novel approach can be used to quantify changes in cell shape following receptor activation and therefore represents a possible additional tool for drug discovery.

Lensless On-chip Imaging of Cells Provides a New Tool for High-throughput Cell-Biology and Medical Diagnostics

Onur Mudanyali¹, Anthony Erlinger¹, Sungkyu Seo¹, Ting-Wei Su¹, Derek Tseng¹, Aydogan Ozcan^{1, 2
1}Electrical Engineering Department, University of California, Los Angeles, ²California NanoSystems Institute, University of California, Los Angeles

Lensfree on-chip imaging and characterization of cells is illustrated. This on-chip cell imaging approach provides a compact and cost-effective tool for medical diagnostics and high-throughput cell biology applications, making it especially suitable for resource poor settings.

Lensless Fluorescent Microscopy on a Chip

Ahmet F. Coskun, Ting-Wei Su, Ikbal Sencan, Aydogan Ozcan
Department of Electrical Engineering, University of California, Los Angeles

A lensless on-chip fluorescent microscopy platform is demonstrated that can image fluorescent objects over an ultra-wide field-of-view of e.g., >0.6-8 cm2 with <4μm resolution using a compressive sampling based decoding algorithm. Such a compact and wide-field fluorescent on-chip imaging modality could be valuable for high-throughput cytometry, rare-cell research and microarray-analysis.