ICCE Equipment - Leica TCS SP8 HCS A

Confocal SystemsLeica TCS SP8 HCS A

Room: S05 V04 F33

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)
  • Photoactivation / Photoconversion
  • automated imaging & screening (Leica HCS A)

Funding

Leica TCS SP8 HCS A microscope was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 219183055.

 

Technical Specifications

Microscope and Accessories

  • inverted Leica DMi8 CS
  • Leica Adaptive Focus Control (AFC)
  • 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
  • Water Immersion Micro Dispenser

Lasers

  • 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: 633 nm (10 mW)

Scanners

conventional (FOV) scanning system: 1 – 1400 Hz (5 fps @ 512 x 512 px), scan format/resolution from 16 x 16 px up to 8192 x 8192 px, zoom 0.75x – 48x

Detector System

  • 3-channel SP-Detector: 2 photomultiplier tubes (PMTs) and 1 super-sensitive Hybrid detector (HyD)
  • 1 PMT for transmitted light (PMT trans)

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 CS2 20x 0.75 dry
HC PL APO 63x/1.4 CS2 63x 1.4 oil
HC PL APO 63x/1.2 W CORR UVIS CS2 63x 1.2 water

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 X (Leica Microsystems) with software modules:

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