FV4000

[Page in progress] The FV4000 by Evident/Olympus is a point-scanning confocal located in CSRL (University Hospital Coventry & Warwickshire), and is the newer version of the FV3000 at Gibbet Hill. One notable upgrade is a new style of more sensitive detectors. The microscope is ideal for imaging deep into living tissue/organisms due to its long working-distance silicone objectives, sensitive detectors, and the superior optical sectioning of a point scanning confocal compared to a spinning disk microscope. The resonant scanners may offer more gentle imaging compared to galvo scanners, although this is somewhat controversial, best to test it yourself and also let us know what happens.

[Manual to come]

Objectives:

• 4x air NA 0.16 WD 13 mm (UPLXAPO4X)
• 10x air NA 0.4 WD 3.1 mm (UPLXAPO10X)
• 20x air NA 0.8 WD 0.6 mm (UPLXAPO20X)
• 25x silicone NA 0.85 WD 2 mm (LUPLAPO25XS), multi-immersion using silicone gel, silicone oil or water. Can also use silicone gel pads to minimise evaporation during time lapses with imaging multiple positions.
• 30x silicone NA 1.05 WD 0.8 mm (UPLSAPO30XS), with a correction collar.
• 40x oil NA 1.4 WD 0.13 mm (UPLXAPO40XO)
• 60x oil NA 1.42 WD 0.15 mm (UPLXAPO60XO), not currently in the microscope

The detectors are a new style promising improved "sensitivity", signal-to-noise, dynamic range, and a more quantitative readout. See Evident's white paper here. The SilVIR detectors have a fixed gain, unlike conventional PMTs, which combined with the improved dynamic range and sensitivity drastically simplifies using the microscope. Saturation is much less of an issue so avoids issues of adjusting the gain to optimise for both dim and bright areas.

The FV4000 has four detectors in total: two red-shifted detectors best for detecting fluorescence longer than about 520 nm, and two broad-band detectors best for wavelengths shorter than about 520 nm.

Nice guide/tutorial to confocal microscopy here.