Live-Cell Imaging | Nikon’s MicroscopyU

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style="background-image:url(https://www.microscopyu.com/assets/poster-images/_galleryPosterFull2x/live-cell-poster.jpg)"></div> <h1>Live-Cell Imaging</h1> <div class="viewoutline"><img src="/images/outline-button_2x.png" width="20" height="17" title="page navigation" /></div></header> <nav class="revnav"> <ul> <li><a href="#rev">Review Articles</a></li> <li><a href="#tuts">Interactive Tutorials</a></li> <li><a href="#gals">Galleries</a></li> <li><a href="#refs">Literature References</a></li> <li><a href="#footnotes">Contributing Authors</a></li> </ul> </nav> <div class="articledescr" id="intro"><p>An increasing number of investigations are using live-cell imaging techniques to provide critical insight into the fundamental nature of cellular and tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein and synthetic fluorophore technology.</p></div> <section class="articlelist" id="rev"> <h2>Review Articles</h2> <ul> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/maintaining-live-cells-on-the-microscope-stage">Maintaining Live Cells on the Microscope Stage</a></h3> <p>A discussion of pH, atmosphere, temperature, media, osmolarity, buffers, and antibiotics.</p> </li> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/live-cell-imaging-culture-chambers">Live-Cell Imaging Culture Chambers</a></h3> <p>Chambers must offer excellent optical properties while maintaining healthy cells.</p> </li> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/introduction-to-fluorescent-proteins">Introduction to Fluorescent Proteins</a></h3> <p>Genetically-encoded fluorescent probes that are revolutionizing live-cell imaging.</p> </li> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/correcting-focus-drift-in-live-cell-microscopy">Correcting Focus Drift in Live-Cell Microscopy</a></h3> <p>Review software and hardware solutions for combating focus drift in time-lapse imaging.</p> </li> </ul> <ul> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/optical-system-and-detector-requirements-for-live-cell-imaging">Optical System and Detector Requirements for Live-Cell Imaging</a></h3> <p>Considerations are detector sensitivity, speed of image acquisition, and photobleaching.</p> </li> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/the-automatic-microscope">The Automatic Microscope</a></h3> <p>Discussion includes shutters, filter wheels, autofocus, light sources, and stage control.</p> </li> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/fluorescent-protein-imaging-parameters">Imaging Fluorescent Proteins</a></h3> <p>Filter requirements, photobleaching, objective choice, highlighters, and multicolor imaging.</p> </li> <li class="intro"><h3><a href="https://www.microscopyu.com/applications/live-cell-imaging/nikon-perfect-focus-system">Nikon Perfect Focus System (PFS)</a></h3> <p>Explore Nikon's high-speed solution for maintaining stable focus in live-cell imaging.</p> </li> </ul> <hr /> </section> <section class="thumblist tutoriallist" id="tuts"> <h2>Interactive Tutorials</h2> <ul> <li> <a href="https://www.microscopyu.com/tutorials/adjustment-of-objective-correction-collars"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/adjustment-of-correction-collars.jpg" height="80" width="80" /> <h3>Adjustment of Objective Correction Collars</h3></a> <p>Learn how to adjust an objective correction collar to minimize spherical aberration.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/choosing-filter-combinations-for-fluorescent-proteins"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/choosing-filter-combos.jpg" height="80" width="80" /> <h3>Choosing Filter Combinations for Fluorescent Proteins</h3></a> <p>Identification of critical filter parameters for imaging fluorescent proteins.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/choosing-fluorescent-proteins-for-dual-labeling-experiments"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/MatchingProteins.jpg" height="80" width="80" /> <h3>Choosing Fluorescent Proteins for Dual Labeling Experiments</h3></a> <p>Interactive tutorial used to optimize pairing of two fluorescent proteins.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/comparison-of-phase-contrast-and-dic-microscopy"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/Phase-DIC.jpg" height="80" width="80" /> <h3>Comparison of Phase Contrast & DIC Microscopy</h3></a> <p>Examples of the same specimen viewed in either phase contrast or DIC.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/dic-microscope-component-alignment"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/dic-alignment.jpg" height="80" width="80" /> <h3>DIC Microscope Component Alignment</h3></a> <p>Examine conoscopic and orthoscopic viewfields in DIC microscopy.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/arclamp"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/arc-lamp.jpg" height="80" width="80" /> <h3>Focus and Alignment of Mercury and Xenon Arc Lamps</h3></a> <p>Explore alignment and focusing of the arc lamp in a mercury or xenon burner, which simulates how the lamp is adjusted in a real microscope. </p> </li> <li> <a href="https://www.microscopyu.com/tutorials/förster-fluorescence-resonance-energy-transfer-with-fluorescent-proteins"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/FRETwFluor.jpg" height="80" width="80" /> <h3>Förster (Fluorescence) Resonance Energy Transfer with Fluorescent Proteins</h3></a> <p>Use this tool to determine the optimum fluorescent protein pairs for FRET. </p> </li> <li> <a href="https://www.microscopyu.com/tutorials/laser-scanning-confocal-microscopy"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/scanning-confocal.jpg" height="80" width="80" /> <h3>Laser Scanning Confocal Microscopy</h3></a> <p>A virtual microscope tutorial featuring a wide variety of specimens.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/matching-camera-to-microscope-resolution"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/matching-resolution.jpg" height="80" width="80" /> <h3>Matching Camera to Microscope Resolution</h3></a> <p>Vary numerical aperture, magnification, and video coupler size to match camera resolution.</p> </li> </ul> <ul> <li> <a href="https://www.microscopyu.com/tutorials/spectralprofiles"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/matching-fluorescent.jpg" height="80" width="80" /> <h3>Matching Fluorescent Probes with Nikon Fluorescence Filter Blocks</h3></a> <p>Explore the various fluorophores that can be imaged with Nikon filter sets. </p> </li> <li> <a href="https://www.microscopyu.com/tutorials/kohler"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/kohler.jpg" height="80" width="80" /> <h3>Microscope Alignment for Köhler Illumination</h3></a> <p>Learn how to adjust a microscope to examine specimens in Köhler illumination. </p> </li> <li> <a href="https://www.microscopyu.com/tutorials/optical-sectioning-with-de-sénarmont-dic-microscopy"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/de-sernamont.jpg" height="80" width="80" /> <h3>Optical Sectioning with de Sénarmont DIC Microscopy</h3></a> <p>At high numerical apertures, DIC can be used for optical sectioning.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/perfect-focus-offset-system-mechanics"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/PFS-offset.jpg" height="80" width="80" /> <h3>Perfect Focus Offset System Mechanics</h3></a> <p>Learn how the offset lens system enables the operation of the Nikon PFS.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/phase-contrast-microscope-alignment"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/phas-alignment.jpg" height="80" width="80" /> <h3>Phase Contrast Microscope Alignment</h3></a> <p>Learn how to align a phase contrast microscope and examine variations in specimen appearance through the eyepieces (at different magnifications) when the condenser annulus is shifted into and out of alignment with the phase plate in the objective.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/phase-plate-configuration-effects-on-specimen-contrast"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/Phase-plate.jpg" height="80" width="80" /> <h3>Phase Plate Configuration Effects on Specimen Contrast</h3></a> <p>Examine contrast variations induced by altering phase plate absorption properties.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/proximity-focused-image-intensifiers"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/proximity.jpg" height="80" width="80" /> <h3>Proximity-Focused Image Intensifiers</h3></a> <p>Light amplification with a micro-channel plate and photocathode.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/spatial-resolution-in-digital-imaging"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/spatial-resolution.jpg" height="80" width="80" /> <h3>Spatial Resolution in Digital Imaging</h3></a> <p>Spatial resolution refers to the number of pixels utilized in construction of the image.</p> </li> <li> <a href="https://www.microscopyu.com/tutorials/the-nikon-perfect-focus-system-pfs"> <img src="https://www.microscopyu.com/assets/thumbnails/_galleryThumbnailSquare/PFS.jpg" height="80" width="80" /> <h3>The Nikon Perfect Focus System (PFS)</h3></a> <p>Examine how the Nikon PFS system operates to maintain stable focus in live-cell imaging.</p> </li> </ul> <hr /> </section> <section id="gals"> <h2>Galleries</h2> <div class="gallerycollection"> <div class="galleryblurb"> <a href="https://www.microscopyu.com/galleries/cell-motility"> <h3>Cell Motility</h3> <img src="https://www.microscopyu.com/assets/poster-images/_galleryThumbnailSquare/licve-cell-poster.jpg" height="80" width="80" /> </a> <p>Explore cell motility in differential interference contrast (DIC) microscopy.</p> <p class="gallerycontents">Movies</p> </div> <div class="galleryblurb"> <a href="https://www.microscopyu.com/galleries/confocal"> <h3>Confocal Microscopy</h3> <img src="https://www.microscopyu.com/assets/poster-images/_galleryThumbnailSquare/confocal-gallery-poster.jpg" height="80" width="80" /> </a> <p>Enjoy the beauty of autofluorescence in thick sections of animal and plant tissues.</p> <p class="gallerycontents">Movies</p> </div> <div class="galleryblurb"> <a href="https://www.microscopyu.com/galleries/dic-phase-contrast"> <h3>Differential Interference Contrast (DIC)</h3> <img src="https://www.microscopyu.com/assets/poster-images/_galleryThumbnailSquare/DIC-gallery-poster.jpg" height="80" width="80" /> </a> <p>Compare specimen contrast using these complementary imaging techniques.</p> <p class="gallerycontents">Image Comparisons • Movies</p> </div> <div class="galleryblurb"> <a href="https://www.microscopyu.com/galleries/fluorescence"> <h3>Fluorescence Microscopy</h3> <img src="https://www.microscopyu.com/assets/poster-images/galleries/_galleryThumbnailSquare/Fluorescence-gallery-poster.jpg" height="80" width="80" /> </a> <p>Cells and tissues examined with synthetic fluorophores in fluorescence microscopy.</p> <p class="gallerycontents">Images</p> </div> <div class="galleryblurb"> <a href="https://www.microscopyu.com/galleries/phase-contrast"> <h3>Phase Contrast</h3> <img src="https://www.microscopyu.com/assets/poster-images/_galleryThumbnailSquare/Phase-Contrast-gallery-poster.jpg" height="80" width="80" /> </a> <p>Specimen viewfields examined using positive and negative phase contrast.</p> <p class="gallerycontents">Image Comparisons</p> </div> <hr /> </div> </section> <section class="referencelist" id="refs"> <h2>Selected Literature References</h2> <ul> <li><h3><a href="https://www.microscopyu.com/references/live-cell-imaging">Live-Cell Imaging</a></h3> <p>Basic concepts in maintaining and imaging living cells on the microscope stage.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/focus-drift-in-optical-microscopy">Focus Drift in Optical Microscopy</a></h3> <p>Focus drift describes the inability to maintain focus over an extended period of time.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/fluorescent-proteins">Fluorescent Proteins</a></h3> <p>Fluorescent proteins are currently the probes of choice for live-cell imaging.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/fluorescence-recovery-after-photobleaching">Fluorescence Recovery after Photobleaching</a></h3> <p>FRAP is used to determine the mobility of fluorescently tagged proteins in live cells.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/chromophore-assisted-light-inactivation">Chromophore-Assisted Light Inactivation</a></h3> <p>CALI is a useful technique for selectively deactivating target molecules.</p> </li> </ul> <ul> <li><h3><a href="https://www.microscopyu.com/references/live-cell-imaging-chambers">Live-Cell Imaging Chambers</a></h3> <p>Chambers specially designed to keep cells alive and healthy on the microscope stage.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/electron-multiplying-ccd-cameras">Electron-Multiplying CCD Cameras</a></h3> <p>EMCCDs are the detector system of choice for live-cell imaging in real-time applications.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/fluorescent-protein-biosensors">Fluorescent Protein Biosensors</a></h3> <p>Genetically-encoded FRET biosensors are useful in detecting a many biological events.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/synthetic-fluorophores-and-hybrid-tags">Synthetic Fluorophores and Hybrid Tags</a></h3> <p>A number of fluorophores have been developed for imaging living and fixed cells.</p> </li> <li><h3><a href="https://www.microscopyu.com/references/fret-microscopy-with-spectral-imaging">FRET Microscopy with Spectral Imaging</a></h3> <p>Excellent for ratiometric analysis of photobleaching and spectral imaging FRET.</p> </li> </ul> <hr /> </section> <div id="footnotes"> <h2>Contributing Authors</h2> <p><b>Michael E. Dailey</b> - Department of Biological Sciences and Neuroscience Program, 369 Biology Building, University of Iowa, Iowa City, Iowa, 52242.</p> <p><b>Daniel C. Focht</b> - Bioptechs Inc., 3560 Beck Road, Butler, Pennsylvania, 16002.</p> <p><b>Alexey Khodjakov</b> and <b>Conly L. Rieder</b> - Wadsworth Center, New York State Department of Health, Albany, New York, 12201, and Marine Biological Laboratory, Woods Hole, Massachusetts, 02543.</p> <p><b>George H. Patterson</b> and <b>Jennifer Lippincott-Schwartz</b> - Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, 20892.</p> <p><b>David W. Piston</b> - Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, 37232.</p> <p><b>Melpomeni Platani</b> - Gene Expression Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117, Heidelberg, Germany.</p> <p><b>Kenneth R. Spring</b> - Scientific Consultant, Lusby, Maryland, 20657.</p> <p><b>Jason R. Swedlow</b> and <b>Paul D. Andrews</b> - Division of Gene Regulation and Expression, MSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland.</p> <p><b>Yu-li Wang</b> - University of Massachusetts Medical School, 377 Plantation Street, Suite 327, Worcester, Massachusetts, 01605.</p> <p><b>Jennifer C. Waters</b> - Nikon Imaging Center, LHRRB Room 113C, Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts, 02115.</p> <p><b>Joel S. Silfies</b> and <b>Edward G. Lieser</b> - Nikon Instruments, Inc., 1300 Walt Whitman Road, Melville, New York, 11747.</p> <p><b>Tony B. Gines</b>, <b>Alex B. Coker</b>, <b>Nathan S. Claxton</b>, <b>Scott G. Olenych</b>, <b>John D. Griffin</b>, and <b>Michael W. 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