LocationRoom 1.19, 1st floor, MRC Building DescriptionUpright Axioplan II research fluorescence microscope suitable for fluorescence and transmitted light 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. 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 are neutral density (ND filters) filters near the focus control 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. Objective LensesMagnification | Immersion | Designation | NA | WD (mm) |
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5X | Air | Fluar | 0.25 | 12.5 | 10X | Air | EC Plan Neofluar | 0.3 | 5.2 | 25X | Multi | Plan Neofluar | 0.8 | 0.21 | 40X | Oil | Plan Neofluar | 1.3 | 0.2 | 63X | Oil | Plan Apochromat | 1.4 | 0.18 | 100X | Oil | Plan Neofluar | 1.3 | 0.2 |
All lenses are corrected for #1.5 coverslips (0.17 micron thick). NA = Numerical Aperture, WD = Working Distance Fluorescence Filter SetsFilter Set | Excitation Filter | Dichroic Mirror | Emission Filter | Fluorophores |
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Zeiss FS02 | G365 | FT395 | LP420 | DAPI, Hoechst | Zeiss FS10 | BP450-490 | FT510 | BP515-565 | FITC, GFP | Zeiss FS26 | BP575-625 | FT640 | BP660-690 | Cy5, Alexa 594 | Blank | N/A | N/A | N/A | Transmitted Light | Zeiss FS43 | BP545/25 | FT570 | BP605/70 | Cy3, TRITC |
You can calibrate the images in both software packages so the number of nanometres per pixel is applied to the data, but note that the Axioplan microscope stand has a set of Optovar lenses that can be used to change the magnification of the image. You must make sure the optovar magnification matches the calibration applied in the software otherwise the calibration factor will be wrong. Only 1x and 1.6x magnifications are supported in the acquisition software. |