Fluorescence platform

 

Responsable Plateau: This email address is being protected from spambots. You need JavaScript enabled to view it.

Contact scientifique: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

The plateau comprises 2 axes:

 

1- Fluorescence spectroscopy

This equipment is dedicated to fluorescence and luminescence characterization, of biomacromolecular interactions or folding, enzymatic activity, protein expression... either from gels, solutions dispatched in multi-well plates, cells on petri dishes or multi-well plates. Some readers can do kinetics measurements, with or without injection.

Please click on specific equipment to get full description.

Typhoon FLA 9500 Fluorescence scanner for gels
Berthold Mithras LB940  
Microplate reader for Absorption, Fuorescence and Luminescence
TECAN safire2 Microplate reader for Fluorescence polarization / anisotropy measurements
CLARIOstar Microplate reader for absorption, fluorescence and polarization measurements
Fast kinetics
Synergy H1 Fluorescence reader for microplates
Cytation 3 Cell imaging in microplates
Amersham Imager 600 ECL reader for Western Blots

 

2- Advanced microscopy and fluorescence spectroscopy

 

The microscopy plateau gathers advanced, custom built setups for various microscopy techniques. Most of this equipment is operated by research engineers and projects are run on the basis of scientific collaborations.  FBI logo final Haute resolution 

Some of these steups belong to MARS facility.

FCS 2 photons FCS, FCCS, N&B and RICS. Laser scanning.
smFRET TCSPC Confocal-based smFRET, FCS, FCCS, PIE, TCSPC, MFD
  mars logo
2 photons STED WorkInProgress 50px

FCS, FCCS, N&B, RICS...
3D STED, fastFLIM, STED FCS
possibility to do FCCS without crosstalk (PIE)
Laser scanning

PALM PALM, STORM, SPT-PALM, TIRF
TIRF / MFM TIRF with dualview on one side => SPT, smFRET, with Alex
3D-MFM on the other side => Multi-focal microscopy
Equipped with microfluidics pump.
Zeiss Axio Imager Upright fluorescence microscope
Rheometer FCS WorkInProgress 50px Blood cell diffusion under shear stress. 2-foci FCS

 


 Back to top

Typhoon FLA 9500 Z2-7@CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications


Laser scanner for fluorescence imaging => precise quantitation of proteins, nucleic acids, and other biomolecules.

Fluorescence detection on gels, membranes, petri dishes, multi-well plates...

 TyphoonFLA9500

 

Specifications

  • High resolution and quantitation: a pixel resolution of up to 10 µm and a linear signal response over five orders of magnitude provides precise quantitation in gels, blots, tissue sections, and arrays
  • High sample throughput: scanning area of 40 × 46 cm enables simultaneous imaging of up to 20 mini-gels or blots
  • 3 lasers : 473, 532, 635nm + 4 emission filters :
    • BPB1 (520-540nm)
    • BPG1 (560-580nm)
    • LPG (≥ 575nm)
    • LPR (≥ 665nm)

Comments

High sensitivity.

Get more information @ the manufacturer website:
https://www.gelifesciences.com/en/us/shop/typhoon-fla-9500-p-00190

 


  Back to top

 Berthold Mithras LB940 Z2-7 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Main applications:
Luminescence, BRET.

Other applications:
FRET, fluorescence, absorption.

Specifications

Microplates from 6 to 1536 wells.
Shaking
Temperature control (heating)
Endpoint or kinetic measurements
Built-in reagent injectors

Mostly used for luminescence detection

Comments

For absorption and Fluorescence, less performant than Tecan or CLARIOstar.

Get more information @ the manufacturer website:
https://www.berthold-bio.com/microplate-readers/details/product/mithras-lb-940/action/show.html

BertholdMithrasLB9940 

 


  Back to top

TECAN safire2 Z2-7 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Mostly polarization / anisotropy => molecular interactions measurements
Microplate reader.

Specifications

Fluorescence reader for microplates
UV-Vis absorbance
Fluorescence intensity
Luminescence
Fluorescence polarization
Temperature control (heating only)
Shaking
Endpoint or kinetic spectral measurements
Possibility to run excitation / emission spectra matrices

Absorption / Fluorescence
Double monochromators in excitation and emission
bandwidth of 10nm in UV, 20nm in visible light
wavelength from 230nm to 850nm

Polarization
4 LEDs : 488, 532, 594, 633nm + double monochromators in detection

Comments

The absorption / fluorescence lamp decays in intensity and cannot be replaced => it is better to use the CLARIOstar for these applications.
Polarization detection is highly sensitive.

TecanSafire 

 

 


 

 Back to top

 CLARIOstar Z2-7 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Absorption / Fluorescence / Polarization measurements.
Fast kinetics.
Multinanodrop in fluorescence. Using the L-vis plate => 2ul volumes, 16 samples simultaneously.


Specifications

Wavelength selection by variable filters for excitation and emission
Instantaneous absorption spectrum (detection on a diodes strip) => super fast kinetics
Fluorescence polarization at all wavelengths


Comments

Possibility to add injectors to get even faster kinetics.
Fluorescence polarization at all wavelengths, contrary to the TECAN.

https://www.bmglabtech.com/clariostar-plus/
ClarioStar 

 

 

 


  Back to top

 Synergy H1 Z2-7 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Fluorescence reader for microplates
UV-Vis absorbance
Fluorescence intensity
Luminescence
Fluorescence polarization
Time-resolved fluorescence

Specifications

Variable filters
Endpoint, or kinetic measurements
Spectral scanning, well area scanning
6- to 384-well plates

Comments

Essentially used to run long kinetics (O/N)
Get more information @ the manufacturer's website:
https://www.biotek.com/products/detection-hybrid-technology-multi-mode-microplate-readers/synergy-h1-hybrid-multi-mode-reader/overview/

 
SynergyH1 

 

 


 Back to top

 Cytation 3 Z2-7 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Fluorescence reader in microplates

Specifications

  • 6- to 384-well plates, microscope slides
  • double monochromators in excitation and emission.
  • Live cell assays with temperature control up to 45°C
  • Endpoint or time-lapse assays
  • Dual reagent injectors

 

Cytation3 

 

 


 

 Back to top

 Amersham Imager 600 Z2-7 @CBS 29 Contact : This email address is being protected from spambots. You need JavaScript enabled to view it.

 


Applications

ECL reader for Western Blots.

Specifications

  • Highly sensitive CCD camera (has to be refrigerated to -20°C)
  • ECL imaging
  • Epi-light source
  • Generation of a combined color image of a chemiluminescence sample image and a white light image of a colored marker.
  • Automatic, manual or semi-automatic exposure mode.
  • Built in fast analysis software. Intuitive design, easy to use image acquisition and analysis workflow.
  • Data can be exported and analysed with Amersham software or any other image analysis software (ImageJ for instance).

Comments

Not connected to the server => use USB port for data transfer. 

 



 


 

 Back to top

 FCS 2photons Z2-17 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

FCS, FCCS, N&B, RICS...
Simultaneous acquisition in 2 emission channels.

Specifications

  • Mai Tai Laser 700-1020nm. 2 photons excitation
  • ISS fast laser scanning module
  • Detection on 2 low dark counts APDs with 5 different dichroic and filters sets to detect fluorescence from classical dyes (CFP, GFP, mCherry, Cy5...).
  • Epi-illumination lamp
  • Nikon statif + 60X NA 1.4 oil
  • 60X NA 1.2 water (=> deeper in the sample + less chromatic aberrations)
  • z-piezo on the objective
  • no auto-focus system.
  • Different sample holders
  • No heating system, no CO2 control.

 

FCS2Photons 

 

 


  Back to top

 smFRET TCSPC Z3-13 @CBS 29 This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Confocal-based smFRET, FCS, FCCS, PIE, TCSPC, MFD

Specifications

Pulsed white supercontinuum source (100ps / 50ns) split in two optical paths, one of them is delayed (7m => 21ns delay)
Band-pass filters to select excitation wavelengths (488, 532, 590, 633, 740nm).
Prompt and delayed beams are recombined in a single mode optical fiber (to clean the beam + ensure proper overlay of all wavelengths)
Pulse synchronizer on the prompt path APM-400, B&H
Nikon statif with perfect focus system (cannot be used at 740nm excitation).
Objectives : 100X NA 1 .4 oil
60X NA 1.2 water
Sample motorized stage (MS-2000, ASI)
Dual polarisation + dual color emission splitter => 2+2 APD (MFD detection)
TCSPC counting board (SPC-150, Becker&Hickl) + 4 channel router

Comments

For full description, please refer to: Olofsson et al, Opt Expr (2013)
https://www.osapublishing.org/oe/abstract.cfm?uri=oe-21-3-3370


 
smFRET TCSPC 

 

 


  Back to top

 2photons / STED                      
mars logo 70px@MARS

This email address is being protected from spambots. You need JavaScript enabled to view it.

This email address is being protected from spambots. You need JavaScript enabled to view it.

This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

FCS, FCCS, N&B, RICS...
3D STED, fastFLIM, STED FCS
possibility to do FCCS without crosstalk (PIE)

Specifications

Pulsed laser Mai Tai 680-1050nm
ISS laser scanning module with 2APD + 2PMT
Detection of classical dyes (CFP, GFP, mCherry, Cy5).
Epi-illumination lamp
Nikon statif + 60X NA 1.4 oil
60X NA 1.2 water (=> deeper in the sample + less chromatic aberrations)
z-piezo on the objective + used to be an autofocus, should be reinstalled.
Sample motorized stage (MS-2000, ASI)
Fast FLIM module

4 Picoquant lasers (405, 485, 595, 640nm) + combiner
Picoquant SepiaII controller => individual laser control for PIE, slaved to the Mai Tai master clock
Shape of the STED laser by an SLM (double reflection on the SLM => lateral then axial PSF engineering).

Comments

This machine will require MRI login.


 

2Photons STED 

WorkInProgress 15th

 

 


 

 Back to top

 PALM mars logo 70px@MARS

This email address is being protected from spambots. You need JavaScript enabled to view it.

This email address is being protected from spambots. You need JavaScript enabled to view it.

 

 

Applications

PALM, STORM, SPT-PALM, TIRF

Specifications

Custom-made inverted microscope (Zeiss stage)
objective: Plan-Apocromat, 100x, 1.4NA oil DIC (Zeiss)
z-axis piezoelectric stage (P-721.CDQ, PICOF, PI).
1.5x telescope (pixel size ≈105 nm)
Lasers:
405 nm (OBIS, LX 405-50, Coherent Inc.)
488 nm (OBIS, LX 488-50, Coherent Inc.)
561 nm (Sapphire)
640 nm (OBIS, LX 640-100, Coherent Inc.)
AOTF: AOTFnc-400.650-TN, AA (opto-electronics)
Emission filters: ET525/50 m, ET600/50 m and ET700/75m (Chroma Technology)
EMCCD camera (iXon X3 DU-897, Andor Technologies)
Sample motorized stage
Home-made auto- focus
Camera, lasers and filter wheel are controlled with software written in Labview.

Comments

This machine requires MRI login.
For full description, please refer to: Georgieva et al, Methods (2016)
https://www.sciencedirect.com/science/article/pii/S1046202316300639

 

 

 


 

 Back to top

 TIRF / 3D-MFM mars logo 70px@MARS This email address is being protected from spambots. You need JavaScript enabled to view it.
This email address is being protected from spambots. You need JavaScript enabled to view it.

Applications

TIRF with dualview on one side => SPT, smFRET, with Alex
3D-MFM on the other side => Multi-focal microscopy

Specifications

Inverted microscope (Zeiss Axiovert 200) in wide field configuration.

MFM setup :

Lasers :                                                                                                   

561 nm (Sapphire 561-100 CW, Coherent, UK)
405 nm (Stradus 405-100, Vortran, USA)
AOTF (AOTFnC-400.650-TN AAoptics, France)
First relay lense L1 (f = 150 mm(to form a secondary Fourier plane, conjugate of the objective pupil plane)
Custom-built multifocus grating (MFG)
Chromatic correction grating (CCG)
Multifacet prism, combined with a second relay lens L2 (f = 200mm)
emCCD camera (IXON DU-897, 512*512, 16µm, Andor, Ireland)
cylindrical lens in front of the em-CCD (for astigmatism).

TIRF setup:
450, 488, 532, 561, 635nm lasers
opto-split => dual view for smFRET or dual color tracking
emCCD camera (IXON 888, Andor, 1024*1024, 13µm)

Comments

This machine will require MRI login.
For full description of the 3D-MFM, please refer to: Oudjedi et al, Biomed Opt Expr (2016)
https://www-ncbi-nlm-nih-gov.fr/pmc/articles/PMC4918573/


 
WorkInProgress 50px 

 

 


  Back to top

 2-foci FCS / Rheometer  @MARS This email address is being protected from spambots. You need JavaScript enabled to view it.
This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Blood cell diffusion under shear stress.

Specifications

633nm laser
2 foci
4-cores fiber => split the signal from the two foci to 2 APDs

Comments

This machine will require MRI login.





 

WorkInProgress 50px 

  Back to top

 Zeiss Axio Imager.A1
@MARS This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Applications

Upright fluorescence microscope

Specifications

Zeis camera
Lumencor Light engine (je sais pas pourquoi la lampe halo a été remplacée par le lumencor)
Objectives :
10x / 0.25
20x / 0.22 DIC
50x / 0.55 DIC




 


 

 

 

 

 

Connexion