Trouble Shooting: Tissue Outline Shifts

STEP 1. 2. 3. 4. 5. 6.

1: Grid Tune Current Lens (using Stage Micrometer) Insert S12 Stage Micrometer onto stage and focus Start with lowest objective first (1.25x) Make sure objective matches the objective selected on the screen. Set reference point close to micrometer. Select Tools >> Grid Tune Current Lens Set Box Size to 300 m (circumference of circle is 600 m - only the circle visible on 100x)

7. Drag icon of anchor to the outer left middle point of circle. 8. Drag dotted lines as needed (entire circle should consist of two white boxes) 9. Right click when finished and click on Finish Calibrating Current Lens 10.Repeat for all objectives. STEP 1. 2. 3. 4. 5. 6. 2: Paracentric/Parafocal Calibration Start with highest objective first (100x). Use putty to hold the slide in place (place putty on the top right corner of slide and stage) Find the micrometer under oil immersion before initiating calibration. Select Paracentric/Parafocal Calibration under Tools Select Video only >> OKAY Select 63x, 40x, 5x and Discard Lenses from Calibration. Select OK.

7. Press OK. Start with the 100x objective and Press OK again.

8. Select a spot on micrometer that you will use for all magnifications (I prefer the + right above the micrometer)

9. After you click on the spot under 100x, the following box will appear:

10.Carefully wipe the oil away from the slide with lens tissue (do not use Kimwipe) 11.Switch objectives to 20x. 12.Change the exposure under Tools >> Adjust Camera Settings and refocus 13.Click on the same spot under 20x:

14.After you click on the calibration spot on 20x, the next prompt will appear:

15.Repeat the same steps for 10x, 2.5x, 1.25x. When finished, select Enable Parcentric and Enable Parfocal.

STEP 3: TEST STAGE MOTOR AND REFERENCE POINT: 1. At the lowest objective (be sure same objective is selected in program), select reference point close to object. 2. Trace the object.

3. Right Click >> Joy Track

4. Turbo move the camera to a completely different spot on the stage. The outline will move with the joystick.

5. Right Click >> End Joystick Mode (outline will now disappear) 6. Move >> To reference Point

7. Camera should now move back to original reference point with the outline still on the object.

8. Repeat for all objectives. As objectives are switched, remember to select the correct objective on the program. Adjust camera settings and exposure as needed. After all 3 calibration steps have been performed, there should be very little outline shift as the objectives are switched. There may be about 10-20 m shift between outlining on 1.25x and switching to 20x. Do not draw your outline exactly on the edges of the tissues, leave a little space between tissue and outline. After calibration, recheck using the stage micrometer.

After all 3 calibration steps have been performed, there should be very little outline shift as the objectives are switched. There may be about 10-20 m shift from outlining on 1.25x and switching

to 20x. Do not draw your outline exactly on the edges of tissues, leave a little space between tissues and outline. After calibration, recheck using stage micrometer.