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Training

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Location

Room 1.19, 1st floor, MRC Building

DescriptionDescription

Location:

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. 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 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.

Objective Lenses

Magnification

Immersion

Designation

NA

WD (mm)

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 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

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.

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