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Russ College of Engineering and Technology, Ohio University

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Generation and Labeling of Murine Bone Marrow-derived Dendritic Cells with Qdot Nanocrystals for Tracking Studies

Maria Muccioli^*1, Michelle Pate^*2, Omowaleola Omosebi², Fabian Benencia^1,2,3

¹Molecular and Cell Biology Program, Ohio University, ²Department of Biomedical Sciences, College of Osteopathic Medicine, Ohio University, ³Department of Biomedical Engineering, Russ College of Engineering and Technology, Ohio University

Dendritic cells uptake antigens and migrate towards immune organs to present processed antigens to T cells. Qdot nanocrystal labeling provides a long-lasting and stable fluorescent signal. This allows tracking of dendritic cells to different organs by fluorescent microscopy.

Bioengineering

Simultaneously Capturing Real-time Images in Two Emission Channels Using a Dual Camera Emission Splitting System: Applications to Cell Adhesion

Grady E. Carlson¹, Eric W. Martin², Monica M. Burdick^1,2

¹Department of Chemical and Biomolecular Engineering, Russ College of Engineering and Technology, Ohio University, ²Biomedical Engineering Program, Ohio University

Dual camera emission splitting systems for two-color fluorescence microscopy generate real-time image sequences with exceptional optical and temporal resolution, a requirement of certain live cell assays including parallel plate flow chamber adhesion assays. When software is employed to merge images from simultaneously acquired emission channels, pseudocolored image sequences are produced.

Bioengineering

Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow

Monica M. Burdick^1,2, Nathan M. Reynolds^1,2, Eric W. Martin², Jacquelyn V. Hawes¹, Grady E. Carlson¹, Chaz M. Cuckler¹, Michael C. Bates¹, Steven R. Barthel³, Charles J. Dimitroff³

¹Department of Chemical and Biomolecular Engineering, Ohio University, ²Biomedical Engineering Program, Russ College of Engineering and Technology, Ohio University, ³Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School

Compared with traditional affinity chromatography using protein A agarose bead-packed columns, protein A membrane adsorbers can significantly speed laboratory-scale isolation of antibodies and other Fc fragment-expressing proteins. Appropriate analysis and quantification methods can further accelerate protein processing, allowing isolation/characterization to be completed in one workday, instead of 20+ work hours.