optical Protocols

Category: Microscopy Protocols: Live-Cell Imaging Protocols

This protocol describes a sealed preparation that allows the continuous long-term observation of cultured mammalian cells on upright or inverted microscopes without environmental CO2 control. The preparation allows for optical conditions consistent with high-quality imaging and good cell viability for at least 100 hours. - [Read A Sealed Preparation for Long-Term Observations of Cultured Cells]

Category: Microscopy Protocols: Confocal Microscopy Protocols

Basic information on confocal microscopy, includes: Specimen Preparation and Imaging; Objective Lens Parameters and Optical Section Thickness; The Objective Lens; Probes for Confocal Imaging; Autofluorescence; Collecting Images; Troubleshooting; Image Processing and Publication; - [Read Confocal Microscopy: Speciman Preparation and Imaging]

Category: Microscopy Protocols: Optical Microscopy Protocols

When imaging specimens in the optical microscope, differences in intensity and/or color create image contrast, which allows individual features and details of the specimen to become visible. Contrast is defined as the difference in light intensity between the image and the adjacent background relative to the overall background intensity. In general, a minimum contrast value of 0.02 (2 percent) is needed by the human eye to distinguish differences between the image and its background. - [Read Contrast in Optical Microscopy]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Specimen chambers have had many designs published over the years describing systems that offer excellent optical properties while allowing specimens to be maintained for varying amounts of time. Ranging in complexity from the simple preparation of a sealed coverslip on a microscope slide to sophisticated perfusion chambers that enable tight control of virtually all environmental variables culture chambers are designed to to allow living specimens to be observed with minimal invasion at high res. - [Read Culture Chambers for Live-Cell Imaging]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Live-cell imaging techniques provide critical insight into the fundamental nature of cellular & tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein & synthetic fluorophore technology. Because of these advances, live-cell imaging has become a requisite analytical tool in most cell biology labs. Includes: Maintaining Live Cells on the Microscope Stage; Live-Cell Imaging Culture Chambers; Optical System and Detector Requirements etc. - [Read Introduction to Live-Cell Imaging Techniques]

Category: Molecular Imaging: Light Microscopy

Light Microscopy - Microscopes in Cell Biology. House Ear Institute. Fluorescence microscopy, Nomarski differential interference contrast, Comparison between phase contrast and interference contrast optical systems , Interference contrast, Phase contrast, Darkfield illumination, alignment of Kohler illumination system, Protocol for using oil immersion lenses, Use of immersion oil, Calculating the final magnification on the photomicrograph, vibration, The coverslip glass, Photomicroscopy. - [Read Light Microscopy - Microscopes in Cell Biology]

Category: C. elegans Protocols

At low magnification, dissection microscopes are useful, whereas at higher magnification and resolution, a wide range of optical methods can be used, including differential interference contrast (DIC) optics, phase contrast, fluorescence, polarized light, confocal, and dark field. - [Read Live Imaging of Caenorhabditis elegans: Observation of Nematodes and Data Collection Protocol]

Category: Microscopy Protocols: Confocal Microscopy Protocols

Confocal laser scanning microscopy (CLSM) is a relatively new light microscopical imaging technique which has found wide applications in the biological sciences. The primary value of the CLSM to the biologist is its ability to produce optical sections through a 3-D specimen-e.g., an entire cell or a piece of tissue - that, to a good approximation, contain information from only one focal plane. Article includes principle and applications of confocal laser scanning microscope. - [Read Looking Inside Cells and Tissues by Optical Sectioning with a Confocal Laser Scanning Microscope]

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Multiphoton fluorescence microscopy is a powerful new technology that enables the acquisition of optical sections without the use of a pinhole aperture typically used for confocal microscopy. The technique is based upon the two-photon principle: A fluorescent molecule simultaneously absorbs two photons producing an electronic transition from the ground to excited state equal to two times the energy of each incident photon. - [Read Multiphoton Images from LSM 510 NLO System]

Category: Microscopy Protocols: Optical Microscopy Protocols

Diffraction-limited optical microscopy requires that the spatial resolution of an image is limited by the wavelength of the incident light & by the numerical apertures of the condenser & objective lens systems.The development of near-field scanning optical microscopy (scanning near-field optical microscopy) has allowed for a imaging technique that retains the various contrast mechanisms afforded by optical microscopy methods while attaining spatial resolution beyond the optical diffraction limit - [Read Near-Field Scanning Optical Microscopy]

Category: Microscopy Protocols: Optical Microscopy Protocols

Near-field scanning optical microscopy can achieve spatial resolution performance beyond the classical diffraction limit by employing a sub-wavelength light source or detector positioned in close proximity to a specimen. Such a sub-wavelength source usually consists of an aperture at the end of a tapered probe, which functions basically as a wave guide. Includes info.: Fiber Probe Fabrication; Pulling Method; Meniscus Etching; Selective Etching; Apertureless and Alternative Probe Designs etc. - [Read Near-Field Scanning Optical Microscopy: NSOM Probes]

Category: General Research Protocols and Methods: Spectrophotometry Protocols

Spectrophotometric Measurement of Nucleic Acids' Concentration Tool. This bioinformatic program help calculate the concentration of nucleic acids according to optical density (including DNA, RNA, oligonucleotides). Zbio - [Read Spectrophotometric Measurement of Nucleic Acids' Concentration Tool]

Category: Microscopy Protocols: Optical Microscopy Protocols

TIRM is a optical technique for monitoring the instantaneous separation distance between a microscopic sphere & a flat plate. Changes in distance as small as 1 nm can be detected. Includes information on: Scattering Intensity I is Related to Elevation h ; apparatus. - [Read Total Internal Reflection Microscopy]