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Zeiss 200M Axiovert Inverted Widefield/Confocal
Light Microscope

This light microscope features a10X conventional and 20X DIC objectives and 40X, 63X, and 100X DIC oil-immersion objectives.

Mercury arc lamps are used for fluorescence microscopy instead of lasers. This gives a wide choice of fluorophores, and is not limited to 2 or 3 specific excitation lines, as with lasers. Also, Hg illumination allows the excitation of UV and short wavelenght blues dyes, such as DAPI and the Hoechst nuclear dyes. Excitation at UV wavelengths with laser-scanning confocal (LSC) microscopes requires either expensive UV-lasers or special multi-photon microscopes. The BBPIC microscope as well has 340 nm and 380 nm excitation filter cubes in the reflector turret for calcium ratioing studies.

Confocal microscopy is done using a spinning Nipkow disc. With the addition of the Andor iXon electron multiplying CCD (EMCCD) camera the BBPIC offers clients confocal imaging without the necessity of a bright signal. This provides the potential of imaging living cells and other samples that are sensitive to the phototoxicity and photobleaching of laser based systems. The iXon will allow confocal imaging in real time at a maxium of 10,000 frames per second.

Image series can be collected as Z-stacks through a sample, as a series of time-lapse images, or both, as time-lapse Z-stacks. These series may also be acquired at a single-wavelength, or as multiple-wavelength images for multiple labeling studies.

Specimens may also be examined by transmitted brightfield illumination, transmitted-light differential interference contrast (DIC), asymmetric illumination contrast (AIC), or epi-fluorescence.

Images collected by any of these methods may then be deconvolved using the microscope's software. The software and hardware are fully integrated so that the imaging program acquires the information needed to do the deconvolution when the image is acquired.

Image recording is done with a cooled CCD monochrome digital camera. Image files may be saved in the Zeiss format or in JPG, TIF, or other formats.
This is a service instrument for use by any group that needs it. The microscope can be operated by BBPIC staff, or investigators may be trained for independent use.