Cell Adhesion & Migration
ISSN: 1933-6918 (Print) 1933-6926 (Online) Journal homepage: http://www.tandfonline.com/loi/kcam20
Live-cell techniques—Advances and challenges Claire M Brown To cite this article: Claire M Brown (2014) Live-cell techniques—Advances and challenges, Cell Adhesion & Migration, 8:5, 429-429, DOI: 10.1080/19336918.2014.996469 To link to this article: http://dx.doi.org/10.1080/19336918.2014.996469
Published online: 20 Jan 2015.
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Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=kcam20 Download by: [Inter-American Development Bank (KNL/FHL)]
Date: 06 November 2015, At: 10:53
FOREWORD Cell Adhesion & Migration 8:5, 429; September/October 2014; © 2014 Taylor & Francis Group, LLC
Live-cell techniques—Advances and challenges Claire M Brown
Downloaded by [Inter-American Development Bank (KNL/FHL)] at 10:53 06 November 2015
McGill University Life Sciences Complex Advanced BioImaging Facility (ABIF) and Department of Physiology, McGill University
The continued development of new imaging modalities and probes has allowed the field of live-cell imaging to blossom. Whether a single molecule, a single cell, cells within tissue or an entire intact organism we are just beginning to explore the mysteries of life in action. Most cells are never exposed to light during their entire existence. So, during light microscopy experiments care must be taken to minimize light exposure and phototoxicity is essential. If the conditions for live-cell imaging are not optimized one may end up studying how cells respond to light induced stress rather than the actual physiology processes of interest. In this special focus issue on Live-cell techniques, Richard Cole provides a review entitled “Live-cell imaging “The cell’s perspective”. He outlines the factors that need to be considered when performing live-cell imaging experiments and how to control them. One sure way to minimize photo-toxicity is to work with brightfield or transmitted light techniques rather than fluorescence. James Jonkman, Pina Colarusso and their research team present the tried-and-true wound healing assay that is widely used to characterize cell migration. They present this seemingly straightforward technique and highlight many of the unforeseen challenges in performing accurate and reproducible wound healing measurements. They discuss the
assay in the context of the overall issue of standardization and reproducibility in science which has recently been a central issue of discussion in the scientific community (Nature Special Issue: Challenges in irreproducible research. 515, 2014; Blow, N.S. A Simple Question of Reproducibility, BioTechniques, 56(1):8, 2014). Moving live-cell imaging experiments from 2D cell culture models to more physiologically relevant 3D environments is an important step forward. Donna Webb’s group presents a research paper where they have moved their work on the Rho family GEF, Asef2, from cells growing in 2D on matrix coated coverslips to 3D collagen matrix model systems. They use novel microfluidic devices to generate reproducible 3D collage matrices and demonstrate that Asef2 regulates cell migration in 3D though a Rac, myosin II dependent mechanism. When live-cell techniques are performed the multidimensional data sets that are generated are rich in quantitative information. To fully harness the plethora of information within these data sets and to decipher the molecular mechanisms at play in biological processes requires the development of models that can be tested using systems biology approaches. Jay Unruh, Brian Slaughter and colleagues present a review article where they use the yeast Cdc42 actin-based polarity mechanism as a case
study to present quantitative metrics for protein interactions, protein quantification and protein dynamics. Their review touches on advanced biophysical imaging techniques such as fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS) as well as reproducibility and standards necessary for calibrated and accurate quantitative live-cell imaging. The special focus issue concludes with the work of Spencer Shorte and his team on the development of a novel super-resolution imaging technique using conical diffraction illumination, termed biaxial super-resolution (BSR). BSR has the hallmarks required for live-cell applications; that is speed and low phototoxicty. In addition, no special sample preparation is required, standard fluorophores can be used and the techniques is compatible with most objective lenses. They have designed BSR as an affordable upgrade to a commercial confocal laser scanning microscope. BSR will open up the possibilities of live-cell techniques to see the molecular processes in the cell with ever more detail and have more precisely localized quantitative information. Overall, this special focus issue demonstrates the care that must be taken when performing live-cell techniques and the wide array of possibilities these techniques open up for scientific investigation.
http://dx.doi.org/10.1080/19336918.2014.996469
www.landesbioscience.com
Cell Adhesion & Migration
429
ISSN: 1933-6918 (Print) 1933-6926 (Online) Journal homepage: http://www.tandfonline.com/loi/kcam20
Live-cell techniques—Advances and challenges Claire M Brown To cite this article: Claire M Brown (2014) Live-cell techniques—Advances and challenges, Cell Adhesion & Migration, 8:5, 429-429, DOI: 10.1080/19336918.2014.996469 To link to this article: http://dx.doi.org/10.1080/19336918.2014.996469
Published online: 20 Jan 2015.
Submit your article to this journal
Article views: 36
View related articles
View Crossmark data
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=kcam20 Download by: [Inter-American Development Bank (KNL/FHL)]
Date: 06 November 2015, At: 10:53
FOREWORD Cell Adhesion & Migration 8:5, 429; September/October 2014; © 2014 Taylor & Francis Group, LLC
Live-cell techniques—Advances and challenges Claire M Brown
Downloaded by [Inter-American Development Bank (KNL/FHL)] at 10:53 06 November 2015
McGill University Life Sciences Complex Advanced BioImaging Facility (ABIF) and Department of Physiology, McGill University
The continued development of new imaging modalities and probes has allowed the field of live-cell imaging to blossom. Whether a single molecule, a single cell, cells within tissue or an entire intact organism we are just beginning to explore the mysteries of life in action. Most cells are never exposed to light during their entire existence. So, during light microscopy experiments care must be taken to minimize light exposure and phototoxicity is essential. If the conditions for live-cell imaging are not optimized one may end up studying how cells respond to light induced stress rather than the actual physiology processes of interest. In this special focus issue on Live-cell techniques, Richard Cole provides a review entitled “Live-cell imaging “The cell’s perspective”. He outlines the factors that need to be considered when performing live-cell imaging experiments and how to control them. One sure way to minimize photo-toxicity is to work with brightfield or transmitted light techniques rather than fluorescence. James Jonkman, Pina Colarusso and their research team present the tried-and-true wound healing assay that is widely used to characterize cell migration. They present this seemingly straightforward technique and highlight many of the unforeseen challenges in performing accurate and reproducible wound healing measurements. They discuss the
assay in the context of the overall issue of standardization and reproducibility in science which has recently been a central issue of discussion in the scientific community (Nature Special Issue: Challenges in irreproducible research. 515, 2014; Blow, N.S. A Simple Question of Reproducibility, BioTechniques, 56(1):8, 2014). Moving live-cell imaging experiments from 2D cell culture models to more physiologically relevant 3D environments is an important step forward. Donna Webb’s group presents a research paper where they have moved their work on the Rho family GEF, Asef2, from cells growing in 2D on matrix coated coverslips to 3D collagen matrix model systems. They use novel microfluidic devices to generate reproducible 3D collage matrices and demonstrate that Asef2 regulates cell migration in 3D though a Rac, myosin II dependent mechanism. When live-cell techniques are performed the multidimensional data sets that are generated are rich in quantitative information. To fully harness the plethora of information within these data sets and to decipher the molecular mechanisms at play in biological processes requires the development of models that can be tested using systems biology approaches. Jay Unruh, Brian Slaughter and colleagues present a review article where they use the yeast Cdc42 actin-based polarity mechanism as a case
study to present quantitative metrics for protein interactions, protein quantification and protein dynamics. Their review touches on advanced biophysical imaging techniques such as fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS) as well as reproducibility and standards necessary for calibrated and accurate quantitative live-cell imaging. The special focus issue concludes with the work of Spencer Shorte and his team on the development of a novel super-resolution imaging technique using conical diffraction illumination, termed biaxial super-resolution (BSR). BSR has the hallmarks required for live-cell applications; that is speed and low phototoxicty. In addition, no special sample preparation is required, standard fluorophores can be used and the techniques is compatible with most objective lenses. They have designed BSR as an affordable upgrade to a commercial confocal laser scanning microscope. BSR will open up the possibilities of live-cell techniques to see the molecular processes in the cell with ever more detail and have more precisely localized quantitative information. Overall, this special focus issue demonstrates the care that must be taken when performing live-cell techniques and the wide array of possibilities these techniques open up for scientific investigation.
http://dx.doi.org/10.1080/19336918.2014.996469
www.landesbioscience.com
Cell Adhesion & Migration
429
