photographing with the microscope

1. Switch on the microscope with the right hand transformer with the voltage turned to the lowest setting and then slowly increase it as necessary for a visually comfortable image.
   
2. The condenser top should be unscrewed and removed and the condenser itself should be lowered when you are using objectives of less magnification than 10x. For all other objectives the condenser top should remain on and the whole condenser should be near the upper limit of its range.
   
3. The field diaphragm and the aperture diaphragm (in the condenser) should be opened and the rotating condenser turret should show J opposite its right side index line.
   
4. Search your slide using a lower magnification objective and increase magnification as necessary. If you use the 100x lens use immersion oil to join the lens to the slide. No other lens may at any time come in contact with oil. To oil the lens put a small drop on the lens and a small drop on the slide. Then rotate the lens so that the two drops merge.
   
5. When you have the wanted field of view, focus the image of your material, close the field diaphragm until it is visible through the binocular eyepieces, focus its image with the condenser drive and center it using the two large adjustment screws on the condenser carrier. Open the diaphragm again until it just vanishes from view.
   
        
   
6. For brightfield observation remove one eyepiece and close the aperture diaphragm (in the condenser) until it darkens or occludes the outer quarter of the bright circle which is visible down the tube. These steps establish Köhler illumination, first described by Dr. August Köhler over 100 years ago.
   
   For phase contrast observation rotate the condenser turret so that 2 is opposite the index mark on the right side of the condenser when using the 16x Ph and 40x Ph objectives or 3 is opposite the index when using the 100x Ph objective. Replace one eyepiece with the phase telescope (which is focused by sliding the top half of it in and out in relation to the other half). The positive phase ring should now be superimposed on the negative ring by using the two adjustments on the condenser, the one nearest you for east-west movement and the one on the left for north-south movement. Another page has more about contrast.
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7. When you are ready to photograph pull the tubehead beamsplitter to the right, diverting light from the binocular tube to the camera tube and the Nikon Coolpix 990 or the SPOT RT KE camera. Do your final focusing and composing on the video monitor and expose with the remote release.
   
8. If you are using the SPOT camera turn on the camera control unit and launch the SPOT software by clicking its desktop icon. If you don't turn on the camera first you will get an error message when you launch the SPOT software.
   
9. Under Setup/Image Setup you will find a dialogue box which allows you to choose between full colour RGB (24 bit) or black & white (8 bit). It also allows changes to be made to a number of other camera parameters, but please don't fiddle with them or you may cause great grief for other users.

   

10. Next, hit the Live button on the top left of the screen and from the Live Image window hit the Controls . . . button to launch the live image control panel which will look like this:
    
    
    
    
11. Focus your specimen and then adjust the Exposure Brightness slider till you have a well exposed image and adjust the Gamma till the contrast looks good with full detail in both the highlight and the shadow areas of your image. Checking the Fit to Window box will force the whole image into the window so you can make sure what you need is included in the field of view. Unchecking it allows you to see part of the image at real size which makes focusing easier. If you don't need the whole field of view, check Selection rather than Full Chip and draw a box that includes only what you need.
    
    
    
12. Briefly move your prep so that the camera sees only the empty field and press the rainbow button (forth from left) on the toolbar to automatically set the white balance for your lighting. Then press Snap to record the image. After every four or five images take a moment to save your pictures, either as TIFF files if you will be doing measurements or analysis or otherwise as JPEG files. Your images can then be adjusted or made into plates using Adobe PhotoShop. The files can be sent via File Transfer Protocol or FTP to your own computer later if they are saved in the C:/Download directory.