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2.2 Kohler Illumination - Microscope Setup |
Setting
up microscope for Kohler Illumination (transmitted light illumination)
- Power Up the Microscope System
as required
- At front of microscope clear upper and lower limits if set
(2)
- Lower objective turret - using coarse focus buttons (3)
- Swing in 10X objective (dry) - using objective changer
buttons (4)
- Crank condenser up - using condenser focus knob (5)
- Place specimen slide on stage holder (6)
- Turn on transmitted light lamp voltage to about 3.5V - incandescent
light wheel (7)
- Swing out transmitted light detector mirror (white line up)
(8)
- Open field diaphragm (9)
- Swing in neutral density filter if desired (10)
- Swing out polarizer (10)
- Set condenser ring to "HF" (for bright field transmitted
illumination) (11)
- Close condenser aperture iris to minimum (12)
- Lower condenser to about 2 mm above slide - using condenser
focus ( 5)
- Set Walliston prism to "H" (for bright field transmitted
illumination) (13)
- Stop off mercury lamp (push rod in) (14)
- Turn dichroic filter wheel to "1" or "4" (no filter)
(15)
- Pull out DIC wave plate (16)
- Push in scan head shutter rod (17)
- Set fine focus steps to
"S3" (coarse) (2)
- Clear upper and lower limit if set
(2)
- Move objective close to slide with coarse focus button control
(3)
- Focus on specimen with coarse focus button controls (3)
- Fine focus with focus wheel (18)
- Set upper limit - important!
- Ensure lower limit is cleared
- it is more efficient to always leave lower limit unset
- Set field iris to minimum
- Focus condenser so that field iris is in best focus
- Center field iris
- Open field iris to just fill the field of view
- Open condenser aperture iris to maximum
- Kohler illumination is now set
- Objectives may now be rotated with button controls
Using 20X, 40X or 100X Objectives
- Setup for Kohler illumination with the 10X objective
- Upper limit must be set using 10X objective focused on your specimen
- Use objective change buttons to rotate in desired objective
- Set focus steps to "S1" (step on front panel
of microscope)
- Press learn to swing objective to right for oiling
- Ensure objective is unlocked (nose turned counter clockwise) and iris is
fully open (knurled ring rotated clockwise)
- Apply small drop of oil to objective lens (approximately 100 ul)
- Without delay press learn again to swing object back
under specimen
- Set condenser aperture iris to minimum
- Focus on specimen
- If transmitted light or DIC images will be collected refocus and center
condenser
- Condenser aperture iris may be set to maximum
Setting up for DIC (Differential Interference Contrast) with 40X or 100X
objectives only
- Setup for Kohler illumination
- Swing in polarizer
- Push in DIC wave plate (Walliston prism and condenser ring
should be on HF or H, i.e. off)
- Open condenser aperture iris fully
- Adjust polarizer to give the darkest background
- Set condenser ring position to correspond to chosen objective (40X or
100X)
- Set Walliston prism position to correspond to chosen objective (40X or
100X)
- View specimen and adjusting Walliston prism position for a grayish image
with best shading
Last update 03-07-31 version
1.8
