Light MicroscopyConfocal and superresolution microscopy

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.

Objectives:

10x (air) HCPL Fluotar with 11mm working distace and a numerical aperture of 0.3

20x (air) PL APO CS with 0.59mm working distance and a numerical aperture of 0.7

40x (oil) PL APO CS with 0.24mm working distance and a numerical aperture of 0.7

63x (water) HC PL APO CS2 with 0.3mm working distance and a numerical aperture of 1.3

63x (glycerin) HC PL APO motCORR CS2 with 0.3mm working distance and a numerical aperture of 1.3

100x (oil) HCX PL APO STED with 0.1mm working distance and a numerical aperture of 1.4

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.