Leica SP8 gSTED super-resolution confocal and FLIM
Download our operating protocol.
Technical Details:
A full set of technical specifications can be viewed here.
Microscope:
Leica TCS SP8 fully automated epifluorescence confocal microscope, with AOTF and AOBS, white light laser (WLL2), gated HyD detection, gated STED,
and Fluorescence Lifetime Imaging - SMD SP FLIM.
Confocal Scanner:
Field of View Scanner (FD 22mm, 8192X8192, 3600Hz) and Resonant (FD 13mm, 1024X1024, 12000Hz)
Confocal excitation lines:
Argon Ion (65 mW): 458nm,476nm, 488nm, 496nm, 514nm. Diode (50mW): 405nm.
White light laser WLL2 (1.5mW per line - 8 simultaneous possible): tunable from 470nm-670nm. picosecond pulsed, 78 MHz.
STED depletion laser (1.5W): 592nm.
5 Spectral Confocal Detection Channels (400nm-800nm):
1 internal SMD FLIM PMT, 2 internal PMTs, 2 internal HyDs (gated).
1 Confocal Transmitted Light Channel:
PMT
Super-resolution STED:
gated STED lateral FWHM 50nm, cw STED FWHM 80 nm, lateral. Normal confocal axial resolution.
STED Sample Preparation:
Protocol 1.
2 colour STED sample preparation - Protocol 2.
Fluorescence Lifetime Imaging Microscopy SP FLIM:
TCSPC module PicoHarp 300, Analysis software "SymphoTime" SPT2.
Objectives:
Objective Type | Magn. | Contrast | Numerical Aperture | Working Distance (mm) | Immersion |
---|---|---|---|---|---|
HCPL FLUOTAR | 10 | - | 0.3 | 11 | air |
PL APO CS | 20 | - | 0.7 | 0.59 | air |
HC PL APO CS2 | 40 | - | 1.3 | 0.24 | oil |
HC PL APO motCORR CS2 | 63 | - | 1.2 | 0.3 | water |
HC PL APO CORR CS2 | 63 | - | 1.3 | 0.3 | Glycerin |
HCX PL APO STED | 100 | - | 1.4 | 0.1 | oil |
Filters for LQ-HXP 120 Lamp:
Filtersystem | Excitation | Emission | Fluorophores |
---|---|---|---|
A | BP 340-380 | LP 425 | DAPI |
I3 | BP 450-490 | LP 515 | FITC, Cy2, EGFP |
N2.1 | BP 515-560 | LP 590 | TRITC, Cy3, Alexa 546, DsRED |
Software: Leica LAS AF 3: Dye finder, Live Data Mode, MicroLab (FRAP, FLIP, FRET), 3D Visualization, FLIM wizard.
Live Cell Imaging Accessories: Ludin Cube and BOX, with active CO2 control.