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Align Optical Components

Koehler Illumination

PROCEDURE: Transmitted Light
1. Focus on specimen with the objective 

to be used for data collection.
2. Close down the luminous field dia-

phragm (1) while viewing.
3. Raise or Lower condenser vertical 

control slightly (2) until luminous field 

diaphragm image is in focus.
4. Center luminous field diaphragm im-

age using condenser centering screws 

(3).
5. Open luminous field diaphragm (1) to 

edge of field.

6. Adjust contrast using condenser 

aperture diaphragm slider (4). In most 

cases adjust aperture diaphragm slider 

to mark matching the NA of the objec-

tive selected. For 20x, 40x, 63x and 100x 

use step 7.
7. Remove eyepiece (5) and check to see 

that 75%-80% of visible aperture is filled 

with light (more light=better resolution 

but less contrast).
Note: this should be done every time an 

objective is changed.

Benefits of Aligning Optics:

• Evenly illuminated image.
• Brilliant image without reflection or 

glare.
• Minimum heating of specimen.

Summary of Contents for AxioImager A1

Page 1: ...Instructions for Zeiss AxioImager A1 Microscope for Stereology Basic operation instructions as of 10 1 2009 The Stereology Workstation By Jon Ekman...

Page 2: ...fluorescence viewing Press the RL Key Set Reflector Turret to 2 Eyes DUAL 470EX 560EX 515EM 620EM Adjust the Incident Iris Stop Slider until light intensity is adequate Find Focus preferably on a dye...

Page 3: ...crews 3 5 Open luminous field diaphragm 1 to edge of field 6 Adjust contrast using condenser aperture diaphragm slider 4 In most cases adjust aperture diaphragm slider to mark matching the NA of the o...

Page 4: ...oscope and in the software Align for Koehler Illumination Grid Tune Calibration 8 Select Tools Grid Tune Current Lens 9 Tell the software the box size Large grid 250mm Small grid 25mm 10 Drop the anch...

Page 5: ...o ing a pilot study will ensure that quality data is generated by obtaining the best estimates of total cell number Step 1 Set up the subject 1 Number of sections to count De fault is 1 new sections c...

Page 6: ...eing counted NOTE Section Thickness measurements Guard Zones Step 8 Define Dissector Options Set Guard zones so that an over or under estimation of cell totals does not occur Do not count cells that f...

Page 7: ...of Results The first items to appear in the results window are the parameters for your study Marker will show the estimated total cell numbers Do not be alarmed if some of the results are 0 Here is t...

Page 8: ...with setting exposure Gain and other camera specific options 6 Adjust Exposure and white balance using More button in the Camera Set tings window another window will pop up with the basic Olympus Pic...

Page 9: ...3 Plan Apochromat excellent quality cor rect for spherical aberration and corrected for chromatic aberration in red blue and green wavelengths 4 EC Epiplan Enhanced Contrast Univer sal objectives for...

Page 10: ...as the Z drive in the windows environment ITG is equipped with gigabit ethernet and saving to the Z drive is relatively fast Also this permits easy access to the data from any computer in the world u...

Page 11: ...hers 1998 Michalet X Kapanidis A N Laurence T Pinaud F Doose S Pflughoefft M and Weiss S The power and prospects of fluorescence microscopies and spectrosco pies Annu Rev Biophys Biomolec Struct 32 16...

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