Leica TCS SP8 Multi-Photon (MP) Microscope with FLIM


 

Technical Details:

 

 

Microscope:

Leica TCS SP8 fully automated epifluorescence confocal microscope, with AOTF and AOBS, gated HyD detection, multi Photon (MP) and Fluorescence Lifetime Imaging - (FLIM).

On a DM6000 CFS basic frame.

 

 

Filters for EL6000 metal halide Lamp:

Filtersystem

Excitation

Emission

Fluorophores

I3

BP 450-490

LP 515

FITC, eGFP, Cy2

N2.1

BP 515-560

LP 590

TRIC, Cy3, Alexa 546 DsRED

 

Confocal Scanner:

up to 8 laserlines with AOBS and VISIR scanoptic with scanrotation. 12kHZ tandemscanner (confocal pointscanner/true confocal scanning system) with 3 channel spectral detection consisting of 2PMT and 1 HyD. Detection region 400-800nm.

 

Nonresonant mode:

Speed  1-1800 Hz

X2Y Scanner Design for maximum scanfield

Max. 7 pic. / sec. 512 x 512

Max  84 pic. / sec. 512 x 16

Max. resolution 8192 x 8192 or 64 Mpixel

 

Resonant mode:

Linefrequency: 24000 Hz (bidirectional)

Frame rate 512x512 pixel (full screen) up to 40 pic. / sec.

Frame rate bei 512x16 pixel bis zu 428 pic. / sec.

Scan format max.: 832 x 832 pixel

Scan format min.: 16 x 16 pixel

 

Objectives:

Objective Type

Magnification

Numerical Aperture

Working Distance (mm)

Immersion

HPLC FLUOTAR

5x

0.15

12

air

N2.1HCX IRAPO

25x

0.95W

2.5

water

HC PL IRAPO

40x

1.1 WCORR

0.63

water

 

 

Laser excitation lines:

confocal:

  • Argon Ion (65 mW): 458nm,476nm, 488nm, 496nm, 514nm. 
  • HeNe (10 mW) 633nm
  • DPSS (20mW) 561nm

Multi photon:

  • Titan:Saphir laser:

  Chameleon Vision II tunable from 680-1040nm (in simultaneous mode 760-860nm)

  with >2,8 W power (at 800 nm)

  Pulse width ~140 fs

  • Compact OPO tunable from 1030-1280nm with >500 mW power (at 1100 nm)

  Pulse width ~200 fs

 

 

 

2 non descanned reflectet light detection filter sets with 2 HyD and 2 PMT:

 

 

Fluorescence lifetime imaging mycroscopy SP FLIM:

TCSPC module PicoHarp 300, Analysis software "SymphoTime" SPT2.

 

 

Software:

Leica LAS AF 3: Dye finder, 3-D Visualization, FLIM wizard.