McMaster University

9 articles published in JoVE

Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion

JoVE 50226 5/02/2013

Justin Tong¹, Pouya Rezai², Sangeena Salam¹, P. Ravi Selvaganapathy², Bhagwati P. Gupta¹

¹Department of Biology, McMaster University, ²Department of Mechanical Engineering, McMaster University

A semi-automated micro-electro-fluidic method to induce on-demand locomotion in Caenorhabditis elegans is described. This method is based on the neurophysiologic phenomenon of worms responding to mild electric fields (“electrotaxis”) inside microfluidic channels. Microfluidic electrotaxis serves as a rapid, sensitive, low-cost, and scalable technique to screen for factors affecting neuronal health.

Iterative Optimization of DNA Duplexes for Crystallization of SeqA-DNA Complexes

JoVE 4266 11/01/2012

Yu Seon Chung, Alba Guarné

Department of Biochemistry and Biomedical Sciences, McMaster University

Crystal structure of protein–DNA complexes can provide insight into protein function, mechanism, as well as, the nature of the specific interaction. Here, we report how to optimize the length, sequence and ends of duplex DNA for co-crystallization with Escherichia coli SeqA, a negative regulator of replication initiation.

Processing of Primary Brain Tumor Tissue for Stem Cell Assays and Flow Sorting

JoVE 4111 9/25/2012

Chitra Venugopal, Nicole M. McFarlane, Sara Nolte, Branavan Manoranjan, Sheila K. Singh

Stem Cell and Cancer Research Institute, McMaster University

The identification of brain tumor initiating cells (BTICs), the rare cells within a heterogeneous tumor possessing stem cell properties, provides new insights into human brain tumor pathogenesis. We have refined specific culture conditions to enrich for BTICs, and we routinely use flow cytometry to further enrich these populations. Self-renewal assays and transcript analysis by single cell RT-PCR can subsequently be performed on these isolated cells.

Analysis of Pulmonary Dendritic Cell Maturation and Migration during Allergic Airway Inflammation

JoVE 4014 7/23/2012

Rahul Kushwah¹, Jim Hu²

¹Stem Cell and Cancer Research Institute, McMaster University, Hamilton, ²Physiology and Experimental Medicine Research Program, Hospital for Sick Children, University of Toronto

We describe a strategy to monitor maturation and migration of pulmonary dendritic cells in response to ovalbumin in the setting of ovalbumin induced allergic airway inflammation. This strategy can be modified to assess migration of pulmonary dendritic cells in settings of infection.

Detection of Bacteria Using Fluorogenic DNAzymes

JoVE 3961 5/28/2012

Sergio D. Aguirre¹, M. Monsur Ali¹, Pushpinder Kanda¹, Yingfu Li^1,2

¹Department of Biochemistry and Biomedical Sciences, McMaster University, ²Department of Chemistry and Chemical Biology, McMaster University

We have recently reported a novel approach for generating fluorogenic DNAzyme probes that can be applied to set up a simple, "mix-and-read" fluorescent assay for bacterial detection. These special DNA probes catalyze the cleavage of a chromophore-modified DNA-RNA chimeric substrate in the presence of crude extracellular mixture (CEM) produced by a specific bacterium, thereby translating bacterial detection into fluorescence signal generation. In this report we will describe key experimental procedures where a specific DNAzyme probe denoted "RFD-EC1" is employed for the detection of the model bacterium, Escherichia coli (E. coli).

Investigating Intestinal Inflammation in DSS-induced Model of IBD

JoVE 3678 2/01/2012

Janice J. Kim, Md. Sharif Shajib, Marcus M. Manocha, Waliul I. Khan

Farncombe Family Digestive Health Research Institute, Department of Pathology and Molecular Medicine, McMaster University

Experimental models of inflammatory bowel disease have allowed us to examine the complex innate and adaptive immune responses associated with pathogenesis. Using histological scoring, quantification of pro-inflammatory cytokines and myeloperoxidase activity, one can begin to assess these responses seen in inflammatory bowel disease.

Primary Human Bronchial Epithelial Cells Grown from Explants

JoVE 1789 3/26/2010

Asma Yaghi, Aisha Zaman, Myrna Dolovich

Medicine, Faculty of Health Sciences, McMaster University

Here we describe a detailed method for growing primary human bronchial epithelial cells from explants of human bronchial airway tissue including differentiated growth on an air-liquid interface. This method provides an abundant source of primary cells for investigating the role of the airway epithelium in human lung health and disease.

A Tactile Automated Passive-Finger Stimulator (TAPS)

JoVE 1374 6/03/2009

Daniel Goldreich^1,2, Michael Wong², Ryan M. Peters², Ingrid M. Kanics¹

¹Department of Occupational Therapy, Duquesne University, ²Department of Psychology, Neuroscience and Behaviour, McMaster University

We describe a computer-controlled device for investigating the sense of touch: the Tactile Automated Passive-finger Stimulator (TAPS). We describe the components of TAPS, and show how TAPS is used to administer a two-interval forced-choice tactile grating orientation test.

Culture of myeloid dendritic cells from bone marrow precursors

JoVE 769 7/25/2008

Jeanette Boudreau^1,2, Sandeep Koshy^2,3, Derek Cummings², Yonghong Wan²

¹Medical Sciences Program, McMaster University, ²Centre for Gene Therapeutics, McMaster University, ³Department of Chemical Engineering, University of Waterloo

This video demonstrates the procedure for differentiating myeloid dendritic cells from mouse bone marrow. Isolation of mouse tibia and femur, and processing of bone marrow are demonstrated. Pictures demonstrating cell morphology before and after differentiation, and figures depicting cell phenotype and IL-12 production following maturation using CpG are shown.