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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Listserv: I would like to draw your attention to a live webinar that I will be moderating starting at 9:00 am CDT on Wednesday April 27th on the subject of label-free imaging. The webinar is being hosted by the International Journal of Biochemistry and Cell Biology (IJBCB), and sponsored by PhaseFocus. The speakers will include Martin Humphry from PhaseFocus, Davide Danovi from Kings College London, and Peter O'Toole from the University of York, and it will be followed by a live Q&A. Label-free imaging represents an under-utilized cost-effective and user-friendly approach that has already made significant impact on fields such as cell adhesion, cell migration, cell proliferation, apoptosis and nanomaterial characterization. Several companies offering different types of quantitative label-free imaging platforms have recently emerged including PhaseFocus, Phasics, NanoLive, PhiOptics and CytoViva. These different solutions hold the potential to revolutionize the way we image cells, tissue and materials and I invite you all to tune into the webinar! Thanks Josh Joshua Z. Rappoport PhD Director of the Center for Advanced Microscopy and the Nikon Imaging Center at Northwestern University Northwestern University Feinberg School of Medicine 303 E. Chicago Avenue Chicago, IL 60611 (312) 503-4140 http://cam.facilities.northwestern.edu/ http://nic.feinberg.northwestern.edu/ |
Joshua Zachary Rappoport |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Sorry everyone! Here is the link: http://view6.workcast.net/register?cpak=7302640263452844 It's time for lunch... From: Joshua Zachary Rappoport Sent: Friday, April 15, 2016 12:06 PM To: [hidden email] Subject: Upcoming webinar Dear Listserv: I would like to draw your attention to a live webinar that I will be moderating starting at 9:00 am CDT on Wednesday April 27th on the subject of label-free imaging. The webinar is being hosted by the International Journal of Biochemistry and Cell Biology (IJBCB), and sponsored by PhaseFocus. The speakers will include Martin Humphry from PhaseFocus, Davide Danovi from Kings College London, and Peter O'Toole from the University of York, and it will be followed by a live Q&A. Label-free imaging represents an under-utilized cost-effective and user-friendly approach that has already made significant impact on fields such as cell adhesion, cell migration, cell proliferation, apoptosis and nanomaterial characterization. Several companies offering different types of quantitative label-free imaging platforms have recently emerged including PhaseFocus, Phasics, NanoLive, PhiOptics and CytoViva. These different solutions hold the potential to revolutionize the way we image cells, tissue and materials and I invite you all to tune into the webinar! Thanks Josh Joshua Z. Rappoport PhD Director of the Center for Advanced Microscopy and the Nikon Imaging Center at Northwestern University Northwestern University Feinberg School of Medicine 303 E. Chicago Avenue Chicago, IL 60611 (312) 503-4140 http://cam.facilities.northwestern.edu/ http://nic.feinberg.northwestern.edu/ |
George McNamara |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Quantitative optical microscopy - I also encourage this article http://www.jove.com/video/50988/quantitative-optical-microscopy-measurement-cellular-biophysical Video requires (institutional) subscription - the .M files are open access PDF version is open access at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4162510/pdf/jove-86-50988.pdf abstract We describe the use of a standard optical microscope to perform*quantitative*measurements of mass, volume, and density on cellular specimens through a combination of bright field and differential interference contrast imagery. Two primary approaches are presented: noninterferometric*quantitative**phase*microscopy (NIQPM), to perform measurements of total cell mass and subcellular density distribution, and Hilbert transform differential interference contrast microscopy (HTDIC) to determine volume. NIQPM is based on a simplified model of wave propagation, termed the paraxial approximation, with three underlying assumptions: low numerical aperture (NA) illumination, weak scattering, and weak absorption of light by the specimen. Fortunately, unstained cellular specimens satisfy these assumptions and low NA illumination is easily achieved on commercial microscopes. HTDIC is used to obtain volumetric information from through-focus DIC imagery under high NA illumination conditions. High NA illumination enables enhanced sectioning of the specimen along the optical axis. Hilbert transform processing on the DIC image stacks greatly enhances edge detection algorithms for localization of the specimen borders in three dimensions by separating the gray values of the specimen intensity from those of the background. The primary advantages of NIQPM and HTDIC lay in their technological accessibility using "off-the-shelf" microscopes. There are two basic limitations of these methods: slow z-stack acquisition time on commercial scopes currently abrogates the investigation of phenomena faster than 1 frame/minute, and secondly, diffraction effects restrict the utility of NIQPM and HTDIC to objects from 0.2 up to 10 (NIQPM) and 20 (HTDIC) μm in diameter, respectively. Hence, the specimen and its associated time dynamics of interest must meet certain size and temporal constraints to enable the use of these methods. Excitingly, most fixed cellular specimens are readily investigated with these methods. PMID: 24747818 Also IATIA (now Ultima Capital) http://www.ultimacapital.net/iatiaimaging/Publications/Iatia%20Imaging/applicationNotes/comparisonWithOpticalPhaseContrastModalities.pdf Yet another company, Ovizio http://www.ovizio.com/ looks especially useful for cell bioreactors (i-Line), does have a camera for a microscope. and a classic (but not trivial to implement interferometry on a research microscope), Brown AF, Dunn GA 1989 Microinterferometry of the movement of dry matter in fibroblasts. J Cell Sci 92: 379-389. We describe the use of interferometric microscopy coupled with a novel application of Sénarmont compensation for detecting and quantifying the distribution of*dry*matter in cultured cells. In conjunction with video techniques and digital image processing, a two-dimensional, calibrated map of the*dry**mass*distribution in an isolated cell can be obtained and digitally recorded. We have called the technique Digitally Recorded Interferometric Microscopy with Analyser Shift (DRIMAS). The method greatly facilitates the automatic recognition of cells by computer. Recorded time-lapse sequences can be used to establish a database of the growth and motility of specific cells in given experimental conditions. Databases of this type can be analysed to reveal the patterns of growth and locomotory behaviour of individual cells. We describe a systematic method of obtaining parameters of cell size, shape, spreading, intracellular motility and translocation. Auto-correlations and cross-correlations between these parameters can be detected and quantified using time series analysis, revealing potential cause/effect relationships in the mechanisms of growth and motility. Besides characterizing the overall pattern of cell behaviour, these data can also yield information about the instantaneous pattern of intracellular motility. We describe the use of finite element analysis to reveal the dynamics of the intracellular transport of*dry*matter. This yields the pattern of the minimum flow of*dry*matter required to account for the changes in its distribution. Most of this flux is not associated with the movement of visible structures and possibly represents the transport of dissociated components of the cytoskeleton. In chick heart fibroblasts, surprisingly high velocities of nearly 2.0 microns s-1 were detected during the period of increased motility following tail detachment. The total kinetic energy associated with the*dry**mass*flux is a single parameter, which characterizes the instantaneous motility of the cell. We found that the kinetic energy of intracellular motility can be several hundred times greater than the kinetic energy of translocation. Kinetic energy may prove to be a very informative single measure of intracellular motility for assessing the effects of malignant transformation, genetic manipulations, and other experimental treatments on the locomotory machinery of the cell. On 4/15/2016 12:50 PM, Joshua Zachary Rappoport wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Sorry everyone! > > Here is the link: http://view6.workcast.net/register?cpak=7302640263452844 > > It's time for lunch... > > From: Joshua Zachary Rappoport > Sent: Friday, April 15, 2016 12:06 PM > To: [hidden email] > Subject: Upcoming webinar > > Dear Listserv: > > I would like to draw your attention to a live webinar that I will be moderating starting at 9:00 am CDT on Wednesday April 27th on the subject of label-free imaging. The webinar is being hosted by the International Journal of Biochemistry and Cell Biology (IJBCB), and sponsored by PhaseFocus. The speakers will include Martin Humphry from PhaseFocus, Davide Danovi from Kings College London, and Peter O'Toole from the University of York, and it will be followed by a live Q&A. > > Label-free imaging represents an under-utilized cost-effective and user-friendly approach that has already made significant impact on fields such as cell adhesion, cell migration, cell proliferation, apoptosis and nanomaterial characterization. Several companies offering different types of quantitative label-free imaging platforms have recently emerged including PhaseFocus, Phasics, NanoLive, PhiOptics and CytoViva. These different solutions hold the potential to revolutionize the way we image cells, tissue and materials and I invite you all to tune into the webinar! > > Thanks > > Josh > > > Joshua Z. Rappoport PhD > > Director of the Center for Advanced Microscopy and the Nikon Imaging Center at Northwestern University > Northwestern University Feinberg School of Medicine > 303 E. Chicago Avenue > Chicago, IL 60611 > (312) 503-4140 > > http://cam.facilities.northwestern.edu/ > http://nic.feinberg.northwestern.edu/ > -- George McNamara, Ph.D. Houston, TX 77054 https://www.linkedin.com/in/georgemcnamara |
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