A5068414716- Cell Image Analysis Techniques
- Advanced Fluorescence Microscopy Techniques
- Cellular Mechanics and Interactions
- Monoclonal and Polyclonal Antibodies Research
- Advanced Biosensing Techniques and Applications
- Cell Adhesion Molecules Research
- Research Data Management Practices
- Scientific Computing and Data Management
- Single-cell and spatial transcriptomics
- Atomic and Molecular Physics
- Genetics, Bioinformatics, and Biomedical Research
- Cellular transport and secretion
- Laser-induced spectroscopy and plasma
- Force Microscopy Techniques and Applications
- Photoacoustic and Ultrasonic Imaging
- Lipid Membrane Structure and Behavior
- Optical Imaging and Spectroscopy Techniques
- Mass Spectrometry Techniques and Applications
- bioluminescence and chemiluminescence research
- Advanced Electron Microscopy Techniques and Applications
- Glycosylation and Glycoproteins Research
- Spectroscopy Techniques in Biomedical and Chemical Research
- Microtubule and mitosis dynamics
- Protein Kinase Regulation and GTPase Signaling
- Molecular Biology Techniques and Applications
McGill University
2016-2025
Developmental Studies Hybridoma Bank
2024
University of Toronto
2024
University of Leicester
2024
Cell Signaling Technology (United States)
2024
Emory University
2024
University at Albany, State University of New York
2015
New York State Department of Health
2012-2015
Wadsworth Center
2012-2015
Glasgow Caledonian University
2014-2015
Imaging of living cells and tissue is now common in many fields the life physical sciences, instrumental revealing a great deal about cellular dynamics function. It crucial when performing such experiments that cell viability at forefront any measurement to ensure physiological biological processes are under investigation not altered way. Many tissues normally exposed light during their cycle, so it important for microscopy applications minimize exposure, which can cause phototoxicity. To...
Neuropathic pain is a chronic debilitating disease that results from nerve damage, persists long after the injury has subsided, and characterized by spontaneous mechanical hypersensitivity. Although loss of inhibitory tone in dorsal horn spinal cord major contributor to neuropathic pain, molecular cellular mechanisms underlying this disinhibition are unclear. Here, we combined pharmacogenetic activation selective ablation approaches mice define contribution parvalbumin (PV)-expressing...
Continuous adhesion formation and disassembly (adhesion turnover) in the protrusions of migrating cells is regulated by unclear mechanisms. We show that p21-activated kinase (PAK)–induced phosphorylation serine 273 paxillin a critical regulator this turnover. Paxillin-S273 dramatically increases migration, protrusion, turnover increasing paxillin–GIT1 binding promoting localization GIT1–PIX–PAK signaling module near leading edge. Mutants interfere with ternary abrogate effects paxillin-S273...
Image correlation microscopy methodology was extended and used to determine retrospectively the density, dynamics interactions of alpha5-integrin in migrating cells. Alpha5-integrin is present submicroscopic clusters containing 3-4 integrins before it discernibly organized. The integrin nascent adhesions, as identified by presence paxillin, approximately 1.4 times more concentrated, 4.5 clustered much less mobile than surrounding regions. Thus, while are throughout cell, they differ...
Summary Raster image correlation spectroscopy (RICS) is a new and novel technique for measuring molecular dynamics concentrations from fluorescence confocal images. The RICS extracts information about images of living cells taken on commercial systems. Here we develop guidelines performing the analysis an analogue laser scanning microscope. Guidelines typical instrument settings, acquisition settings detector characterization are presented. Using appropriate instrument/acquisition...
The confocal fluorescence microscope has become a popular tool for life sciences researchers, primarily because of its ability to remove blur from outside the focal plane image. Several different kinds microscopes have been developed, each with advantages and disadvantages. This article will cover grid confocal, classic laser-scanning (CLSM), resonant scanning-CLSM, spinning-disk microscope. way technique works, best applications is suited for, limitations technique, new developments...
Cell migration is regulated in part by the connection between substratum and actin cytoskeleton. However, very large number of proteins involved this linkage their complex network interactions make it difficult to assess role cell migration. We apply a novel image analysis tool, spatio-temporal correlation spectroscopy (STICS), quantify directed movements adhesion-related protrusions migrating cells. The STICS technique reveals protein dynamics even when densities are low or high, works...
Paxillin is an adaptor molecule involved in the assembly of focal adhesions. Using different fluorescence fluctuation approaches, we established that paxillin-EGFP dynamic on many timescales within cell, ranging from milliseconds to seconds. In cytoplasmic regions, far adhesions, paxillin uniformly distributed and freely diffusing as a monomer, determined by single-point correlation spectroscopy photon-counting histogram analysis. Near dynamics are reduced drastically, presumably due binding...
Fluorescence illumination can cause phototoxicity that negatively affects living samples. This study demonstrates much of the and photobleaching experienced with live-cell fluorescence imaging occurs as a result 'illumination overhead' (IO). when sample is illuminated but emission not being captured by microscope camera. Several technological advancements have been developed, including fast-switching LED lamps transistor-transistor logic (TTL) circuits, to diminish caused IO. These are...
Antibodies are critical reagents to detect and characterize proteins. It is commonly understood that many commercial antibodies do not recognize their intended targets, but information on the scope of problem remains largely anecdotal, as such, feasibility goal at least one potent specific antibody targeting each protein in a proteome cannot be assessed. Focusing for human proteins, we have scaled standardized characterization approach using parental knockout cell lines (Laflamme et al.,...
Antibodies are critical reagents to detect and characterize proteins. It is commonly understood that many commercial antibodies do not recognize their intended targets, but information on the scope of problem remains largely anecdotal, as such, feasibility goal at least one potent specific antibody targeting each protein in a proteome cannot be assessed. Focusing for human proteins, we have scaled standardized characterization approach using parental knockout cell lines (Laflamme et al.,...
Abstract Antibody-based research applications are critical for biological discovery. Yet, there no industry standards to compare the performance of antibodies in various applications. We describe a knockout cell line-based antibody characterization platform, developed, and approved jointly by academic researchers that enables systematic comparison western blot, immunoprecipitation, immunofluorescence procedures. The scalable protocols consist (i) identification appropriate lines studies,...
Abstract In the dynamic landscape of scientific research, imaging core facilities are vital hubs propelling collaboration and innovation at technology development dissemination frontier. Here, we present a collaborative effort led by Global BioImaging (GBI), introducing international recommendations geared towards elevating careers Imaging Scientists in facilities. Despite critical role modern research ecosystems, challenges persist recognising their value, aligning performance metrics...
5'-AMP-activated protein kinase (AMPK) serves as an energy sensor and is at the center of control for a large number metabolic reactions, thereby playing crucial role in Type 2 diabetes other human diseases. AMPK present nucleus cytoplasm; however, mechanisms that regulate intracellular localization are poorly understood. We have now identified several factors distribution AMPK. Environmental stress regulates AMPK, upon recovery from heat shock or oxidant exposure accumulates nuclei. show...
It is important to determine the most effective method of delivering light onto a specimen for minimal induced damage. Assays are presented measure photo-bleaching fluorophores and photo-toxicity living cells under different illumination conditions. Turning off during part experimental time reduced in manner proportional exposure. The rate EGFP was by 9-fold with pulsing on micro-second scale. Similarly, cells, rapid line scanning resulted cell stress as measured mitochondrial potential,...
Imaging technologies are used throughout the life and biomedical sciences to understand mechanisms in biology diagnosis therapy animal human medicine. We present criteria for globally applicable guidelines open image data tools resources rapidly developing fields of biological imaging.