The equipment

The Zeiss LSM 780 inverted confocal microscope, controlled by ZEN software and equipped with a thermostatic chamber, is fully motorized and enables sub-cellular observation of fixed samples or living cells. The microscope is equipped with a spectral strip for simultaneous observation of up to 8 colors, and a high-resolution detector to improve resolution by a factor of 1.7.

Related technologies :

2D imaging

3D imaging

FRAP photoactivation

Ratiometric FRET

Spectral imaging

High-resolution imaging

Available lenses

Lenses	Digital opening	Immersion
X10 Plan Apo	0.45 DF= 2.1 mm	sec
X40 W C-Apo	1.2 DF= 280 µm	water
X40 DIC Plan Apo	1.3 DF= 200 µm	oil
X63 DIC Plan Apo	1.4 DF= 190 µm	oil

Laser sources

– 405nm diode, 30mW

– Argon laser, lines at 458, 488, 514 nm, 25mW

– 561nm diode, 20 mW

– Diode 633 nm, 5mW

Detectors

2 PMTs (multi-alkali coated photocathodes), one of which is cooled

1 PMT transmission

1 cooled 32-channel multiPMT (GaAsP photocathode) for spectral detection.

1 Airyscan detector comprising 32 GaAsP honeycomb detectors

The Leica Sp8 upright confocal microscope, controlled by LAS X software, enables sub-cellular observation of fixed samples or living cells. It is equipped with deconvolution software for enhanced resolution.

Related technologies:

2D imaging

3D imaging

Multiphoton imaging

Available lenses

Lenses	Digital opening	Immersion
X25 PL IRAPO	0.95 DT= 2.5 mm	water
X40 PLAN APO	1.3 DT=240 µm	oil
X63 PLAN APO	1.4 DT = 280 µm	oil

Laser sources

– 405 nm diode, 50 mW

– Argon Multilines at 458, 488, 514 nm, 65 mW

– Diode at 561 nm, 20 mW

– Diode 633 nm, 10 mW

– Chameleon Vision II pre-compensated IR Coherent laser tunable from 680 to 1080 nm. It features (500-550, 565-605) for maximum observation.

Detectors

It is equipped with 2 PMTs and a HyD

This inverted microscope can perform transmitted light and fluorescence times lapses on living cells. It can also be used for TIRF.

Filters

GFP/mRFP EX DBP 480/30 + 565/25. EM DBP 525/31 + 616/57

It is equipped with :

– a thermostatically-controlled, CO2-regulated chamber for studies on living cells

– an XYZ motorized stage

– high-resolution Cool Snap HQ2 camera

– an Evolve EMCCD camera

– a DG4 with Fura2 filter for Calcium detection

Available lenses

Lenses	Digital opening	Immersion
X2.5 Plan-Neofluar	0.075 df=9.5 mm	sec
X5 Plan-Neofluar	0.16 df= 18.5 mm	sec
X10 Ph 1 Plan-Neofluar	0.5 df= 5.2 mm	sec
X20 Ph2 Plan-Neofluar	0.5 df= 2 mm	sec
X40 DIC Plan-Apo	1.3 df= 0.2 mm	oil
X63DIC Plan-Apo	1.4 df= 0,19 mm	oil
X100 DIC Plan-Apo	1.46 df= 0,10 mm	oil

Filter blocks

Singleband HE	Multiband	Multiband
DAPI EM EX G 365BP 445/50	GFP/mRFP	DAPI/GFP/Cy3/Cy5
eGFP EX BP 470/40, EM BP 525/50	CFP/YFP
CFP EX BP 550/25, EM BP 480/40	Fura 2, EX BP 340/30, EM BP 690/50
YFP EX BP 500/25, EM BP 535/30
Cya3 EX BP 436/25, EM BP 605/70
Cya5 EX BP 640/30, EM BP 690/50

Imaris is a 3D/4D reconstruction software package published by Bitplane. It can be used to visualize images taken using photonic microscopy, by volume and surface reconstruction. It is freely available on the Plate-Forme after reservation on the CRI resources site.

Fiji is very comprehensive for image analysis (co-location, stack processing, colorimetry, edge search, segmentation, z projection, overlay, filters, 3D, .AVI .GIF animation creation, 3D deconvolution, …) and allows you to create macros.

Official website: https://imagej.net/Fiji

X-CLARITY ™ systems and reagents standardize, simplify and accelerate every step of the tissue cleaning process. With the CLARITY method, preserved tissue is embedded in a hydrogel matrix and lipids are actively extracted by electrophoresis to create a stable, optically transparent tissue-hydrogel hybrid that is chemically accessible for multiple cycles of antibody labeling and imaging. Native cytoarchitecture remains intact and even endogenous fluorescence proteins are preserved for robust downstream fluorescence imaging.