PerkinElmer Ultraview Vox Confocal
Location: Room 1.12, MRC Building
The Vox is a spinning disc confocal microscope designed for rapid acquisition of 3D and 4D data. The microscope consists of a Nikon TiE inverted stand attached to a confocal scan head with a Yokogawa CSU-X1 spinning disc that uses a patented microlens design to optimise light transmission. The detector is a Hamamatsu C9100-13 EMCCD camera. EMCCD cameras multiply signal on the chip before reading out. This makes them more sensitive but a large pixel size means that resolution is low compared to a CCD in equivalent conditions. A Prior NanoscanZ piezo focus device allows fast collection of image stacks. The TiE microscope is fitted with the Perfect Focus System (PFS), which maintains samples in precise focus indefinitely (most suitable for samples mounted at a glass/aqueous interface, but there are some exceptions). The Vox system also has a Photokinesis Unit (PKU) for spot or ROI illumination in techniques such as FRAP and photoactivation/photoswitching.
- Objective lenses
- Microscope filter sets
- Lasers
- Emission filters
Using the Vox
- Start up and shut down procedure
- Coming Soon - Familiarising yourself with the Vox user interface
- 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 |
There is also a 100x 1.3 NA Plan Fluor oil objective that can be used if the PFS range turns out to be too narrow when using the 100x Plan Apo.
There is a choice of three immersion oils:
Oil | Refractive Index | For use with... | Kept in... |
|---|---|---|---|
Cargille Type HF | 1.518 (at 23ºC) | Oil lenses at room temperature | Tray outside environmental chamber |
Cargille Type 37 | 1.518 (at 37°C) | Oil lenses at 37ºC | Tray inside environmental chamber |
Zeiss Immersol W | 1.334 (at 23ºC) | Water immersion lens only | Tray inside environmental chamber |
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.
Emission Wheel Filters
Turret Position | Emission Filter (nm) | Example Fluorophores |
|---|---|---|
1 | Empty | N/A |
2 | 527(55) | GFP |
3 | 455(80) 615(70) | DAPI, Texas Red |
4 | 485(60) 705(90) | CFP, Cy5 |
5 | 587(125) | YFP |
6 | DIC | Brightfield DIC |
7 | 525(50) 640(120) | GFP, RFP |
8 | 477(45) 575(100) 705(90) | Hoechst, YFP, Cy5 |
9 | Empty | N/A |
10 | Empty | N/A |
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. |