ICCE Equipment - Leica TCS SP5 MP HCS A

Confocal SystemsLeica TCS SP5 MP HCS A

Room: S05 V05 F83

Main Applications

  • confocal (point) laser scanning microscopy, optical sectioning
  • multi-dimensional live cell + fixed specimen imaging
  • multi-position, tile scan / Mosaic
  • FRAP
  • FRET (acceptor photobleaching & sensitized emission)
  • automated imaging & screening (Leica HCS A)
  • 2-photon microscopy

Technical Specifications

Microscope and Accessories

  • inverted Leica DMI 6000B
  • motorized xy-stage
  • SuperZ Galvo
  • external fluorescence lamp LQ-HXP 120 LEJ
  • Halogen lamp for transmitted light
  • controlled environmental conditions for live cell imaging (temperature, CO2)
  • Microtiterplate insert for SuperZ stage

Lasers

confocal
  • Diode Laser: 405 nm (50 mW)
  • Argon Laser: 458 nm (5 mW), 476 nm (5 mW), 488 nm (20 mW), 496 nm (5 mW), 514 nm (20 mW)
  • DPSS Laser: 561 nm (20 mW)
  • HeNe Laser: 594 nm (2.5 mW) and 633 nm (10 mW)
Multiphoton
  • Ti:Sapphire: 710 – 990 nm (> 1.5 W @ 800 nm), Mai Tai BB (Spectra Physics)

Scanners (tandem scanner system)

  • conventional (FOV) Scanner: 1 – 1400 Hz (5 fps @ 512 x 512 px), scan format/resolution from 16 x 16 px up to 8192 x 8192 px, zoom 1x – 64x
  • resonant Scanner (ultra-fast, reduced photodamage): 8000 Hz (25 fps @ 512 x 512 px), scan format/resolution from 16 x 16 px up to 1024 x 1024 px, zoom 1.7x – 32x

Detector System

  • 5-channel SP-Detector: 3 photomultiplier tubes (PMTs) and 2 super-sensitive Hybrid detectors (HyDs)
  • 1 PMT for transmitted light (PMT trans)
  • 2 NDD (non-descanned detectors): PMT RLD1, PMT RLD2

Beam splitting between excitation and emission light is controlled via AOBS.

Objectives

name

magnification

 NA immersion
HC PL Fluotar 10x/0.3 10x 0.3 dry
HC PL APO 20x/0.75 CS 20x 0.75 dry
HCX PL APO 20x/0.70 imm corr Lbd. BL. 20x 0.7 water, oil,glycerol
HCX PL APO 40x/1.25-0.75 Lbd. BL. 40x 1.25 oil
HCX PL APO 63x/1.40-0.60 Lbd. BL 63x 1.4 oil
HCX PL APO 100x/1.46 100x 1.46 oil
HCX PL APO 63x/1.3 glycerol (upon request) 63x 1.3 glycerol

Filter sets for LQ HXP-120

  • Filterset A (Excitation BP 340-380, Dichroic 400, Emission LP 425)
  • Filterset I 3 (Excitation BP 450-490, Dichroic 510, Emission LP 515)
  • Filterset N 2.1 (Excitation BP 515-560, Dichroic 580, Emission LP 590)

 

Software

LAS AF (Leica Microsystems) with software modules:

  • HCS-A Matrix Screener (for automated microscopy, high content screening)
  • LAS MicroLab
    • FRAP wizard
    • FRET wizard (acceptor photobleaching & sensitized emission)
  • LAS Live Data Mode
  • LAS 3D