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

The Leica SP8 is an automated inverted confocal laser scanning microscope allowing simultaneous acquisition of 5 fluorescent channels and 1 transmitted light channel (BF or DIC). Further, the system is outfitted with a very flexible white light laser source. A climate chamber and resonant scanner favor confocal live cell studies.

Location

University Zurich, Gloriastrasse 30, Room GLM F01.

Training Request

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

Technical Specifications

Microscope

inverted confocal microscope (Leica DMi8 CS with AFC, Model SP8)
5 freely selectable emission detection windows
motorized x/y stage
3D capability
time lapse capability

Light Sources and Lasers

Halogen lamp for transmitted light
External fluorescence lamp LQ-HXP 120
Diode Lasers: 405 nm (50mW), 442nm
Argon Laser (65mW): 458 nm, 476 nm, 488 nm, 496 nm, 514 nm
White light laser (WLL): continuous laser output in the range of 470-670nm; AOTF allows to choose up to 8 different wavelengths simultaneously; pulsed laser system with a repetition rate of 80 MHz

Scanners (tandem scanner system)

confocal point-scanning - optical sectioning
zoom function
scan field rotation
Regular scanner (field-of-view scanner): 1 Hz to 1800 Hz
Resonant scanner: 8 kHz

Detectors

5 confocal fluorescence channels: one Photomultipliertubes (PMTs) and four super-sensitive Hybrid detectors (HyD)
one transmission light-channel (PMT-Trans)

Camera System

Hamamatsu Orca Flash 4.0 V3 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

Objectives

Name	Magnification	NA	Immersion	WD (mm)
HC PL FLUOTAR	10x	0.3	Air	11.0
HC PL APO CS2	20x	0.75	IMM	0.68
HC PL APO	40x	0.95	Air	0.17
HC PL APO CS2	63x	1.4	oil	0.1
HC PL APO CS2motCorr	63x	1.2	water	0.3

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

Fluorescence Filters

Filter for Epi-Fluorescence using the Filter turret

Name	Excitation Range	Excitation Filter	Dichroic	Emission Filter
DAPI	UV	BP 350/50	400	BP 460/50
GFP	blue	BP 470/40	495	BP 525/50
RHOD	green	BP 546/10	560	BP 585/40
QUAD-S	UV/blue/green/red	LP (see fast filter wheels)	425; 505; 575; 660	LP (see fast filter wheels)

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)

Literature and Links

Laser Scanning confocal Microscopy

ibiology - Optical Sectioning and Confocal Microscopy

Leica Science Lab

Responsible Persons

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

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

How to acknowledge contributions of the Center for Microscopy

Center for Microscopy and Image Analysis

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CLSM - Leica SP8 inverse (Gloriastrasse)