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Microscope:
Leica SP8 point-scanning confocal system, on a Leica DMI6000B inverted microscope, fully motorized, with Leica 鈥淪uperZ鈥� Galvo stage, four spectral fluorescent light detectors (three PMT, one HyD high sensitivity detector), one transmitted light detector (PMT)
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Objectives:
|
Position |
Objective |
DIC |
|
1 |
10x/0.4 dry HC PL APO 1.x/0.40 CS |
(prism not installed) |
|
2 |
20x/0.7 dry HC PL APO 20x/0.70 CS |
DIC |
|
3 |
40x/0.85 dry HCX PL APO 40X/0.85 CORR CS,0.11- |
DIC |
|
4 |
40x/1.1 water HC PL APO 40x/1.10 W CORR CS2 |
DIC |
|
5 |
40x/1.3 oil HC PL APO 40x/1.30 Oil CS2 |
DIC |
|
6 |
63x/1.4 oil HC PL APO 63x/1.40 OIL CS2 |
DIC |
Confocal mode:
Lasers, no fluorescence emission filters (all spectral detectors):
|
Lasers (all diode) |
Typical fluorophore |
|
405nm, 50mW, DMOD |
DAPI |
|
448nm, 40mW, AOTF |
CFP |
|
488nm, 20mW, AOTF |
AF488, GFP, YFP |
|
552nm, 20mW, AOTF |
TRITC, Cy3 Texas Red, RFP |
|
638nm, 30mW, AOTF |
AF633, Cy5, FarRedFP |
Laser modulation by AOTF (acusto-optic tunable filters), or DMDO (direct modulation of laser intensity)
Wide-field fluorescence:
Light source: Leica EL6000 metal-halide lamp
Fluorescence filter cubes, Leica:
|
Filter set |
Excitation filter |
Dichroic |
Emission filter |
Typical fluorophore |
|
I3 |
BP450-490 |
510 |
LP515 |
AF488, GFP |
|
N2.1 |
BP515-560 |
580 |
LP590 |
TRITC, RFP |
|
A |
BP340-380 |
400 |
LP425 |
DAPI |
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