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.