CLSM - Leica SP8 inverse (IPZ)

SP8_inverse_IPZ

 

 

The Leica SP8 is an automated upright confocal laser scanning microscope allowing simultaneous aquisition of 3 fluorescent channels and 1 transmitted light channel (BF or DIC).

Location

University Zurich, Institute of Parasitology, Winterthurerstrasse 266a Room: PV-10.55. Phone: 044 635 85 99

Training Request

Follow this link to apply for an introduction to the microscope.

Technical Specifications

Light sources and lasers

  • Halogen lamp for transmitted light
  • External fluorescence lamp  LQ-HXP 120
  • Solid state diode lasers: 405 nm (50 mW), 488 nm (20 mW), 552 nm (20 mW) and  638 nm (30 mW)

Scanners

Dual scanning system:

Regular scanner: 10 - 1800 Hz

Resonant scanner: 8000 Hz

Objectives

Name Magnification NA Immersion WD (mm)
HC PL Fluotar 10x 0.3 Air 11
HC PL APO CS2 20x 0.75 IMM 0.66
HC PL APO CS2 40x 1.1 Water 0.65
HCX PL APO 63x 1.2 Water 0.22
HC PL APO CS2 63x 1.4 Oil 0.14
HC PL APO 100x 1.46 Oil 0.09

CS2 objectives: improved color correction, perfect VIS-405; IMM = multi immersion (either water, glycerol or oil)

Detector System

  • Two Hybrid (HyD) detectors
  • one photomultiplier tube (PMT) detector
  • one transmission light channel (PMT-trans)
  • Excitation controlled by AOTF
  • beam splitters: RT 15/85, substrate, TD 488/552/638, DD 488/552 for separation between excitation and emission
  • Accurate emission detection by spectral detectors.

Camera System

  • Hamamatsu Orca Flash 4.0 V2 high sensitivity, digital monochrome scientific cooled sCMOS camera
  • QE > 82% @ 560 nm
  • 2048 x 2048 pixel (6.5 um x 6.5 um)
  • 16 bit dynamics

Fluorescence Filters

Filter for Epi-Fluorescence using the Filter turret

Name Excitation Range Excitation Filter Dichroic Emission Filter
QUAD-S UV/blue/green/red LP (see fast filter wheels) 425; 505; 575; 660

LP (see fast filter wheels

DAPI UV BP 350/50 400 BP 460/50
DsRed green BP 546/11 560 BP 605/75
GFP blue BP 470/40 495 BP 525/50

Filter for Epi-Fluorescence using the QUAD-S in combination with the fast filter wheels

Filter Name Excitation/Emission Range Filters/Bands

DFTC-EX

UV/blue/green/red

BP 350/50, BP 490/20, BP 555/25, BP 645/30, EMPTY

DFTC-EM

UV/blue/green/red

BP 455/50, BP 525/36, BP 605/52, BP 705/72, EMPTY

 

Accessories

  • controlled environmental conditions (CO2, temperature, humidity)
  • Matrix screening software (reservation needed)

Remarks

This microscope acquires images with improved axial resolution compared with more coventional widefield microscopes. In addition to the multicolor images the microscope can acquire large tile scans fully automatically. The microscope allows live imaging with controlled environmental conditions (CO2, temperature, humidity).

Literature and Links

Further information (internal UZH use only)

Follow this link for further background information, documents and links

Responsible Persons

If you have questions about the microscope please contact Jana Döhner or Caroline Aemisegger.