Leica DMI6000B
Room1.035 on the 1st floor of the “Medizinisches Forschungszentrums”Widefield Fluorescence and Time-Lapse Microscope
The Leica DMI6000B is a fully automated epi-fluorescence widefield microscope, optimized for use in live cell microscopy. It has a motorized 3-plate microscope stage that can be moved in x, y and z and autofocus functions (adaptive focus control (AFC) and automatic autofocus).
Transmission illumination is provided by a halogen bulb with a fast-switching shutter in front. Fluorescence illumination is provided by a mercury short-arc lamp. The following excitation wavelengths can be generated via very fast-switching filter wheels:
| 340+/-13nm (e.g. Fura340) | 490+/-10nm (e.g. FITC) |
| 360+/-20nm (e.g. DAPI) | 504+/-6nm (e.g. YFP) |
| 387+/-5nm (e.g. Fura380) | 555+/-12,5nm (e.g. TxRed) |
| 427+/-5nm (e.g. CFP) | 645+/-15nm (e.g. Cy5 oder AF647) |
For optimum fluorescence images, the system has the following automatically swiveling dichroic beam splitters:
| 1. „TRX“ - ECFP / EYFP / mCherry | Tm >90%: 457-483; 522-555; 602-663 |
| 2. „TRS“ - DAPI / FITC / TRITC / (Cy5) | Tm >90%: 437-474; 506-539; 575-622; 665-711 |
| 3. „FU2“ - Fura 2 | Tm >93%: 415-950 |
| 4. „FRE“ - CFP / YFP - FRET | Tm >90%: 449-483; 530-569 |
Images are generated using a Leica K3M CMOS camera (sensor format: 3,072 x 2,048 pixels, pixel size: 2.4 x 2.4 µm). An additional filter wheel with the following configuration is placed in front of the camera, which can be used to filter the emission signals.
| blue: 455+/-25nm | red: 605+/-26nm |
| green: 525+/-18nm | far red: 705+/-36nm |
The following lenses are available in the system:
|
Objective Type |
Magnification |
Contrast Method |
Numerical Aperture |
Working Distance (mm) |
Immersion Medium |
|---|---|---|---|---|---|
|
HC PL FLUOTAR Leica: 506224 |
5x |
BF, DIC, P, DF, IMC, FLUO |
0.15 |
13.7 |
Air |
|
HC PL FLUOTAR L Leica: 506243 |
20x |
BF, DIC, P, DF, PH, IMC, FLUO |
0.4 |
7.7-6.2 |
Air |
|
HC PL FLUOTAR L Leica: 506203 |
40x |
BF, DIC, P, DF, PH, IMC, FLUO |
0.6 |
3.3-1.9 |
Air |
|
HC PL APO Leica: 506220 |
100x |
BF, DIC, DF, FLUO |
1.4 |
0.09 |
Oil |
For use in live cell microscopy, the microscope stand is enclosed in a PECON “Incubator BLX Black” climate chamber. This can be used to incubate and microscope specimens for extended periods at 37 °C and 5 % CO2.
The following LASX software modules are available for the system:
| - 11640032 AF6000 Multi-channel recording |
| - 11640031 AF6000 Time series recording |
| - 11640030 AF6000 z-stack recording and autofocus |
| - 11640029 AF6000 Mark & Find with control of motorized tables |
| - 11640028 AF6000 Stitching (mosaic shots) |
| - 11640025 AF6000 Calcium Imaging |
| - 11640804 AF6000 3D-Visualisation |
>>The microscope can only be used in close consultation with the EMU so as not to disrupt the processes on the other EMU devices!<<