Summary
Here are some highlights from the November 2011 Issue of Journal of Visualized Experiments (JoVE).
Protocol
Measuring Fast Calcium Fluxes in Cardiomyocytes
Urszula Golebiewska1, Suzanne Scarlata2
1Department of Biological Sciences and Geology, Queensborough Community College, 2Department of Physiology & Biophysics, Stony Brook University
We present a method to isolate rapid (microsecond) calcium events from slower fluxes in living cells using laser scanning confocal microscopy. The method measures fluorescence intensity fluctuations of calcium indicators by recording line scans of several hundred pixels in a cell. Histogram analysis allows us to isolate the time scales of different calcium fluxes.
Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets
Helen Tran, Sarah L. Gael, Michael D. Connolly, Ronald N. Zuckermann
Molecular Foundry, Lawrence Berkeley National Laboratory
A simple and general manual peptoid synthesis method involving basic equipment and commercially available reagents is outlined, enabling peptoids to be easily synthesized in most laboratories. The synthesis, purification and characterization of an amphiphilic peptoid 36mer is described, as well as its self-assembly into highly-ordered nanosheets.
Behavioral Assessment of Manual Dexterity in Non-Human Primates
Eric Schmidlin*, Mélanie Kaeser*, Anne- Dominique Gindrat, Julie Savidan, Pauline Chatagny, Simon Badoud, Adjia Hamadjida, Marie-Laure Beaud, Thierry Wannier, Abderraouf Belhaj-Saif, Eric M. Rouiller
Department of Medicine, University of Fribourg
* These authors contributed equally
As manual dexterity is a prerogative mainly of primates, behavioral tasks have been developed in macaque monkeys. Four reach and grasp prehension tasks, measuring hand manipulation ability and force, allow to establish functional recovery after a lesion of the central nervous system and to test the effect of a treatment.
Determining the Phagocytic Activity of Clinical Antibody Samples
Elizabeth G. McAndrew1, Anne-Sophie Dugast1, Anna F. Licht1, Justin R. Eusebio1, Galit Alter1, Margaret E. Ackerman2
1Massachusetts General Hospital, Ragon Institute of MGH, MIT, and Harvard, 2Thayer School of Engineering, Dartmouth College
We present a high-throughput flow cytometric assay to determine the phagocytic activity of antigen-specific antibodies from clinical samples, utilizing fluorescent antigen-coated beads and a monocytic cell line expressing multiple Fc receptors-providing receptor usage and phagocytic activity determinations in a standardized and reproducible fashion for any antigen of interest.
Study of the Actin Cytoskeleton in Live Endothelial Cells Expressing GFP-Actin
Travis M. Doggett, Jerome W. Breslin
Department of Physiology, Louisiana State University Health Sciences Center Microscopic imaging of live endothelial cells expressing GFP-actin allows characterization of dynamic changes in cytoskeletal structures. Unlike techniques that use fixed specimens, this method provides a detailed assessment of temporal changes in the actin cytoskeleton in the same cells before, during, and after various physical, pharmacological, or inflammatory stimuli.
Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-Moving Laboratory Mice
Nikola Cesarovic1, Paulin Jirkof2, Andreas Rettich2, Margarete Arras1
1Division of Surgical Research, University Hospital Zurich, 2Institute of Laboratory Animal Science, University of Zurich
A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, wellbeing and survival rate are also presented.
Electroporation of Craniofacial Mesenchyme
Jacqueline M. Tabler, Karen J. Liu
Department of Craniofacial Development, King's College London
Craniofacial cartilages develop in close contact with other tissues and are difficult to manipulate in live animals. We are using electroporation to deliver molecular tools during growth of the craniofacial skeleton while bypassing early embryonic effects. This approach will allow us to efficiently test candidate molecules in vivo.
Slice Electrophysiological Recording and Optogenetic Stimulation of Adult-Born Neurons in the Olfactory Bulb
Matthew Valley, Sebastian Wagner, Benjamin W. Gallarda, Pierre-Marie Lledo
Laboratory for Perception and Memory, Institut Pasteur and Centre National de la Recherche Scientifique (CNRS)
Adult-born neurons expressing ChR2 can be manipulated in slice electrophysiological preparations in order to examine their contribution towards the function of olfactory neural circuits.
