SPE3 Microscope Information

DMI4000 microscope

The SPE confocal system is attached to a DMI4000 microscope stand. The fluorescence filter turret and objective lens nose-piece are manually rotated but the positions are encoded so the software knows which objective lens is in place. The microscope stand focus in manual so confocal sectioning is achieved via a galvanometer driven stage insert (Z Galvo). Eyepiece or confocal light path selection is automated.

Objective lenses

Leica ACS objective lenses have been designed specifically for SPE confocal microscopes with 405 nm lasers to correct for colour registration errors between the violet and visible lines. They do not have a flat field so Leica recommends that the confocal zoom is set to ≥1.6.

Fluorescence filter sets (for observation down the eyepieces)

These fluorescence filters are only used when you're focusing on your specimen down the eyepieces. The filter turret is automatically rotated to the empty Position 4 (BF) when a confocal scan is initiated.

Positions 4 to 6 are irrelevant for fluorescence.

Lasers and dichroic mirrors

The SPE3 has the following lasers. Note that it differs from the other SPEs in that it has a 561 nm laser instead of a 532 nm laser.

Dichroic mirrors separate the excitation light from the emitted fluorescence. Without dichroic mirrors or other beam splitting technologies your image would be contaminated with laser light.