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If the lenses have been setup to be parfocal then if focus is found on one objective, you kcna change it will be in-focus (or at least close) on another. This is usefule for find your sample.
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### Setting up Parfocality
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# Setting up Parfocality
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The focus was on histological slide for each lense and Z-position recorded
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There is big difference in the focal plane for the lenses, I’ve checked that they are all screwed in completely. Is this range normal?
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| Lens | Z / um |
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|----------|---------|
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| 20X | 2033.75 |
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| 40X ELWD | 3304.38 |
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| 40X | 5265.08 |
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| 60X | 2005.85 |
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| 100X | 2061.97 |
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| 100X | 2055.35 |
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|Mag | NA | | Imm | Pos | Working dist. | Z / um | website
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|-----|-----|----------------------|-----|-----|--- |----------|---------------------------------------------------------------------------------------------------|
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| 20X | 0.45| Plan Fluor ELWD AD1 |Air | 1 | 8.2 – 6.9 | 2033.75 | [link](https://www.nikoninstruments.com/Product-Selectors/Objective-Comparison/\(items\)/i117)|
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| 40X | 0.6 | S Plan Fluor |Air | 2 | 3.6 – 2.8 | 3304.38 |[link]( https://www.nikoninstruments.com/Product-Selectors/Objective-Comparison/\(items\)/i118) |
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| 40X | 1.0 | Plan Apo |Oil | 3 | 0.16 | 5265.08 |[link](https://www.nikoninstruments.com/Products/Optics/Apochromat-Objectives/CFI-Plan-Apochromat-Series/CFI-Plan-Apochromat-Series-Specifications) |
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| 60X | 0.95| Plan Apo |Air | 4 | 0.21 – 0.11 | 2005.85 |[link](https://www.nikoninstruments.com/Product-Selectors/Objective-Comparison/\(items\)/i99) |
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|100X | 1.4 | Plan Apo VC |Oil | 5 | 0.13 | 2061.97 |[link](https://www.nikoninstruments.com/Product-Selectors/Objective-Comparison/\(items\)/i95) |
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|100X | 1.45| Plan Apo 𝝺 |Oil | 6 | 0.13 | 2055.35 |[link](https://www.nikoninstruments.com/Product-Selectors/Objective-Comparison/\(items\)/i101) |
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Following the instructions from page 84 the Nikon Eclipse Ti RCP manual the Auto-Link Function (ALF) was turned off and the ALF memory for each lens set at the focal points.
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With ALF off when the lens is changed the lens is escaped to 500um, the turret rotated, and the lens is return to intial position.
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Following the instructions from page 84 the Nikon Eclipse Ti RCP manual:
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1. the Auto-Link Function (ALF) was turned off the movements tab
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2. the ALF memory for each lens set at the focal points and saved.
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To test the setup I looked at the effect of swapping between lenses with ALF on and off. To illustrate the problem here is what happens when I change from 40X 1.0 (pos 3) to the 60X (pos 4).
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| ALF status |Change to 60X | Z sequence |
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|------------|---------------------|---------------------------------|
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| OFF | side of Ti | 5290 -> 500 -> 5290
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| OFF | RCP | 5290 -> 500 -> 5290
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| OFF | software TiPad | 5290 -> 500 -> 5290 -> 2000
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| ON | side of Ti | 5290 -> 500 -> 5290 -> 2000
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| ON | RCP | 5290 -> 500 -> 5290 -> 2000
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| ON | software TiPad | 5290 -> 500 -> 5290 -> 2000
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#### ALF off sequence
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1. the lens is escaped to 500um
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2. the turret rotated
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3. and the lens is returned to initial position.
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#### ALF on sequence
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1. the lens is escaped to 500um
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2. the turret rotated
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3. and the lens is return to initial position
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4. lens shifted by the difference of the ALF memory Z heights.
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##Issues
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1. When the ALF is disabled from the RCP and the lenses are changed from from the software Ti Pad the ALF correction is being applied. This is not a major issue.
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2. When the ALF is on, the lens is bought to the original Z position, then the corrected by the difference in ALF positions. This results in the lens ramming the sample for certain combinations (eg 40X 1.0 to 60X).
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1. ALF Off on the RCP
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Initial Focus with the 40X Z=5279
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a. Change to 60X with side of Ti Zf=5290
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b. Change from RCP Zf=5290
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c. Change from Software TiPad Z=5290, Zf=2000
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2. ALF On:
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a. 500, 5290, 2000
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