DMI4000 microscope
The SPE confocal system is attached to a DM4000 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 on the Leica DM4000 Microscope
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.
Magnification |
Immersion Medium |
Designation |
Numerical Aperture |
Coverslip Correction |
DIC Prism |
|---|---|---|---|---|---|
10X |
Air |
ACS APO |
0.3 |
None |
C |
20X |
Multi Imm |
ACS APO |
0.6 |
0 - 0.17 |
|
40X |
Oil |
ACS APO |
1.15 |
0.17 |
E |
63X |
Oil |
ACS APO |
1.3 |
0.17 |
E |
Fluorescence Filter Sets on the Leica DMI4000 Microscope
Turret Position |
Filter Set |
Excitation Filter |
Dichroic Mirror |
Emission Filter |
Example Fluorophores |
|---|---|---|---|---|---|
1 |
A |
BP340-380 |
RKP400 |
LP425 |
Hoechst/DAPI |
2 |
I3 |
BP450-490 |
RKP510 |
LP515 |
FITC/GFP |
3 |
N2.1 |
BP515-560 |
RKP580 |
LP590 |
Texas Red |
4 |
BF |
N/A |
N/A |
N/A |
Transmitted Light |
5 |
N/A |
N/A |
N/A |
N/A |
Empty |
6 |
Pol |
N/A |
N/A |
N/A |
Polariser for DIC and polarised light microscopy |
Positions 4 to 6 are not used for fluorescence microscopy.