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Description

Location: Room 1.19, 1st floor, MRC Building

Upright Axioplan II research fluorescence microscope suitable for fluorescence and differential interference contrast (DIC) imaging of samples that have been fixed and mounted on slides. The 25x objective can also be used for water dipping provided the dish is wide enough to accommodate the lens. The fluorescence filters on the microscope can be used for conventional fluorophores in the blue, green, red and far red regions of the spectrum. Typical fluorophores would be DAPI, FITC, GFP, TRITC, Cy3, Alexa 647 and Cy5. CFP cannot be imaged with the filter sets on this microscope. YFP may be visible in the green channel. If you have an unusual dye then please email the light microscopy facility for advice.

In addition to the objective lenses the microscope stand has an optovar turret just below the eyepiece trinocular that can be used to provide extra magnification. There is a 0.63x demagnification lens between the microscope stand and the camera. This gives the widest field of view for the camera chip but reduces the resolution. You can improve the resolution by using the 1.6x optovar, since 0.63x times 1.6 = 1. You must make sure the optovar position matches the calibration applied in the software otherwise the image magnification will be wrong. Only the 1x and 1.6x magnifications are supported in the acquisition software.

There are neutral density (ND) filters near the focus on the right of the stand to reduce the intensity of the halogen lamp if it is too bright. The microscope has a QImaging QIClick greyscale CCD camera for high sensitivity fluorescence imaging. The Apple Mac computer currently has Openlab and Micro-Manager installed for image acquisition.

Specifications

The camera specifications are on a datasheet on the QImaging website (datasheet). The camera has a chip with 1392 x 1040 pixels and the pixel size is 6.45 microns. To work out the resolution you need to divide this number by the total magnification of the image (objective lens, demagnification lens and optovar) - or you can use the calibration factors stored in Openlab and Micro-Manager.