Perkin Elmer Ultraview Vox Confocal
Location: Room 1.12, MRC Building
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- Objective lenses
- Microscope filter sets
- Lasers
- Emission filters
- Start up and shut down procedure
- Further Information
Objective Lenses
Magnification | Immersion Medium | Designation | Numerical Aperture | Coverslip Correction | Working Distance(mm) |
|---|---|---|---|---|---|
10X | Air | Plan Apo | 0.45 | 0.17 | 4 |
20X | Air | CFI Plan Apochromat VC | 0.75 | 0.17 | 1 |
40X | Air | CFI Plan Fluor | 0.75 | 0.17 | 0.72 |
60X | Oil | CFI Plan Apochromat | 1.4 | 0.17 | 0.13 |
100X | Oil | CFI Plan Apochromat VC | 1.4 | 0.17 | 0.13 |
60X WI | Water | CFI Plan Apochromat | 1.2 | 0.17 | 0.27 |
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Note that Cargille Type 37 is optimal for use at 37ºC but introduces spherical aberrations when imaging beyond about 10 μm in aqueous media because it is not refractive indexed matched to water. Zeiss Immersol W in combination with the 60x WI lens might be a better choice in this case but its RI is corrected for use at room temperature and won't be correct at 37 degrees.
Lasers and emission filters
The Vox has diode or DPSS lasers that emit the following wavelengths: 405 nm, 488 nm, 561 nm and 640 nm.
Filter cubes in the microscope stand
The filter cubes below are for use when viewing your specimen down the microscope eyepieces. The filter turret must be in position 1 (EMPTY) when imaging with the spinning disc.
Position Number | Filter Set | Excitation Filter | Dichroic Mirror | Emission Filter | Example Fluorophores |
|---|---|---|---|---|---|
1 | EMPTY | N/A | N/A | N/A | None. For use with bright field or when imaging with the spinning disc |
2 | DAPI | D350/50x | 400DCLP | D460/50m | DAPI/Hoechst/AMCA |
3 | FITC | HQ470/40x | Q495LP | HQ525/50m | Endow GFP |
4 | TRITC | HQ575/50x | Q610LP | HQ640/50m | HcRed1, mCherry |
5 | ANALY | N/A | N/A | N/A | DIC analyser (for viewing DIC in eyepieces) |
6 | EMPTY | N/A | N/A | N/A | None. Do not use. |
Further Information
Setting up the Nikon Perfect Focus System (PFS) for multi-point acquisition in Z
The following is a procedure I've worked out for setting up the PFS for multi-point acquisition. I used fluorescent HeLa cells grown on Mat-Tek glass-bottomed 35 mm dishes and mounted in a Solent Scientific insert designed for a range of 35 mm dishes but adjusted to hold this one firmly in place. I cannot say for sure that the procedures will work for all surfaces. Nikon only recommend glass of standard #1.5 coverslip thickness (170 µm).
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