eTraining Introduction

Specimen Preparation

Leica Ultracut UCT



Hitachi S-4700 FE-SEM

Hitachi FB-2000A FIB


Veeco Dim 3000 AFM

Fluorescence Microscopes


Basic Operation

This section describes live viewing in transmitted light.

AxioVision Toolbar

AxioVision Toolbar

The software and icons refer to the AxioVision software for Windows. Click the toolbar above for a larger view.

1. Use the viewing binocular to locate an area of interest on your sample.    
 1.1. The LCD display will tell you to which source (camera or binocular) the light is directed. VIEW
 1.2. To direct light to the binocular, check the LCD display. If the light is going to the binocular, the display will read: Binocular 100%. If this is not the case, press the LEFT/ RIGHT sideport key on the bottom left of the microscope base twice. VIEW
2. To achieve proper contrast, make sure nothing is blocking the light pathway. You may also need to try different phase plates to obtain the best contrast. VIEW
3. Use the coarse focus knob on the bottom right of the microscope to focus the image. VIEW
Live Icon4. In the AxioVision software, open the AxioCamMR: Live window by selecting the Live icon.    
5. Depress the LEFT/ RIGHT sideport key twice to divert all of the light to the camera. VIEW
6. With each change of the objective lens and sample, you will need to check the alignment of the condenser.

To do so, move the slider on the turret away from you until a hexagon appears in the live viewing window. Use the black condenser adjustment knob to focus the hexagon. Once focused, move the slider towards you until the edges of the hexagon just surpass the viewing window.
7. Refocus the image with the coarse and/or fine adjustment knobs. The camera and binocular are not in the same focal plane, so the images from the two sources will not have the same point of focus.    
8. To change the magnification, press the forward or backward OBJECTIVE keys on the right side of the microscope base to rotate through the objective lenses. The objective lens in use should match the phase plate. VIEW
9. To correctly adjust the exposure time for the image, select Acquisition> Cameras > AxioCamMR. VIEW
10. Use the slider to manually adjust the exposure.

NOTE: In the AxioCamMR: Live window, there are several options available to further assist with exposure adjustment. These options can be manipulated to adjust the exposure, if desired. The most commonly used combination involves the A. Best Fit, G0.45, and Over Exp. options.

Do not select the Auto Snap or Auto Live options.
Snap Icon11. Once you are satisfied with the quality of the image, select the Snap icon in the AxioCamMR: Live window.    
12. Save your images as both a .zvi and a .tif by going to File > Save as > your user directory.

To create a user directory, go to the C: drive > Users folder > add a folder titled with your first and last names.

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