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| The DBC | FACULTY | RESEARCH THEMES | FACILITIES | INFO |
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Zeiss Axiovert 200M motorized inverted microscope is equipped with a Zeiss LSM 510 META multi-spectral analyzer. The entire system is user-controlled from a PC. |
The LSM 510 META is suited for many forms of conventional confocal microscopy. To find additional information about methods of confocal microscopy using LSM510META, objectives, how to separate different spectra of fluorophores, how to get around problems with "bleed-through" of signal and much more, read on, or visit the OBC's confocal microscopy resource page. |
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In addition to the tuneable multi-photon MaiTi laser, this LSM 510 META (named "META2") is equipped to produce the following laser lines 458 nm, 477nm, 488nm, 514nm (all Ar-laser), 543nm (HeNe), 633nm (HeNe). This microsope does not have a multi-photon laser or a UV laser to excite in the UV range. However, excitation of dyes (e.g. DAPI) in the UV range in conjunction with scanning microscopy, can be accomplished using the Xcite lamp as described here. OBC support staff can instruct users how to do this. To determine which wavelength you require to excite your fluorophore, download a pdf containing information about optimal excitation and emission wavelength for commonly used fluorophores, or go to the BD web site to use a Java Applet, or visit the database at the Chroma website. |
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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 (short pass, 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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Dr. Michelle Digman, the Director of the Optical Biology Core Facility, provides hands-on training and after-training support for all microscope users. Please note: due to the complex and sensitive nature of the equipment, all users are required to be certified by the Manager for use of the OBC microscopes before using the facility. Please contact Dr. Digman to arrange a training session. |
| For more information about this system and its capabilities, please contact Dr. Michelle Digman, the Director of the Microscopy and Image Analysis Facility. | |
