CLSM - Leica SP8 inverse (IPZ) | Center for Microscopy and Image Analysis

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.

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
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).

Make sure to acknowledge the Center for Microscopy in your publication to support us.

How to acknowledge contributions of the Center for Microscopy

Literature and Links

iBiology_optical_sectioning_and_confocal_microscopy

Molecular expressions_optical_microscopy_primer

Leica_Science_Lab

Further information (internal UZH use only)

Follow this link for further background information, documents and links

Responsible Persons

If you have questions about the device please contact the responsible person.