Upright fluorescence microscope with DIC
Location
Room 1.19, 1st floor, MRC Building
Description
Upright 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. 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.
Introductory training includes how to align the microscope correctly for transmitted light (Koehler illumination), fluorescence and transmitted light imaging using Openlab, applying calibration to the images, applying colour look-up tables, saving and exporting data.
Objective Lenses
Magnification | Immersion | Designation | NA | Contrast | Coverslip Correction (mm) | WD (mm) |
---|---|---|---|---|---|---|
5X | Air | Fluar | 0.25 | None | 0.17 | 12.5 |
10X | Air | EC Plan Neofluar | 0.3 | None | 0.17 | 5.2 |
25X | Multi | Plan Neofluar | 0.8 | DIC | 0 - 0.17 | 0.21 |
40X | Oil | Plan Neofluar | 1.3 | None | 0.17 | 0.2 |
63X | Oil | Plan Apochromat | 1.4 | None | 0.17 | 0.18 |
100X | Oil | Plan Neofluar | 1.3 | None | 0.17 | 0.2 |
NA = Numerical Aperture, WD = Working Distance
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.
Axioplan Filter Sets
Filter Set | Excitation Filter | Dichroic Mirror | Emission Filter | Fluorophores |
---|---|---|---|---|
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 |