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Zeiss Axiovert 200M motorized inverted microscope is equipped with a Zeiss LSM 510 META multi-spectral analyzer, and a Spectra-Physics MaiTai Ti-Sapphire two-photon femtosecond laser. System is controlled from a PC. This microscope is equipped with DIC (Nomarski) optics. The laser lines, objectives and filter sets (in scan head, and in filter turret on microscope stand) are listed below. This system also has a Pecon XL3 incubator that provides thermal stability to the microscope during time-lapse imaging. In addition, the incubator has an internal chamber in which the humidity and CO2 can be regulated to enable time-lapse imaging of cells. |
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| The LSM 510 META is suited for many forms of conventional confocal microscopy, but is especially suited to perform advanced analyses including Fluorescence Recovery After Photobleaching (FRAP), and Fluorescence Resonance Energy Transfer (FRET). With acquisition of additional software, these experiments can be conducted in an easily quantifiable manner. To find additional information about methods of confocal microscopy using LSM510META, multi-photon lasers, objectives, how to separate different spectra of fluorophores, how to get around problems with "bleed-through" of signal and much more, visit the OBC's confocal microscopy resource page. |
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In conventional epifluorescence microscopy, a filter turret or slider contains a set of three filters (excitation, dichromatic and emission) that select the wavelength of light that is used to illuminate (excite) the specimen and the wavelength of light (emission) permitted to be be viewed through eyepieces or by a camera. In contrast, in confocal microscopy, multi- or single- line lasers that emit light of specific wavelengths (instead of a broad spectrum as you'd get from a mercury arc lamp) are used to illuminate a sample. See above for the wavelengths of light that can be produced by this confocal. With regard to the emission filters, the META detector scanhead contains a series of filters (band pass, long pass) that can be selected electronically via the confocal software. A significant advantage of this scanhead is the META detector itself, which can resolve a variety of wavelengths in 20nm intervals, similar to a spectrophotometer. This enables separation of a wide range of spectra with similar wavelengths. For more information on which emission filter sets are available, check this Table. |
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| For more information about this system and its capabilities, please contact Dr. Michelle Digman, the Director of the Optical Biology Core Facility, or Jenny Sasaki, the Core's Jr. Specialist. |
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