SBRI Stem-cell and Brain Research Institute - France (Lyon)

Technical Facilities : Live Imaging

This core facility includes a 2 photons microscope and two Time Lapse Video inverted microscopes.

Composition of team / September 1, 2017

Coordinator : Veronique Cortay

  • Veronique Cortay - Technician (TR)
  • Dorothée Patti - Engineer (AI)
  • Nathalie Beaujean - DR2
  • Irène Aksoy - CR

Description Equipment

Two-photon microscope

The microscope is an inverted Axio-Observer Z1 from Zeiss, equipped with a Chameleon system Ultra (I) Titane Sapphire 80 Mhz laser. Tuning range wavelength : 690 to 1040 nm.

  • One monophoton laser at 543 nm
  • Software is Zeiss LSM 510.
  • Microscope with motorized objective nosepiece.
  • Plan Apo-Chromat 10x/0.45, without immersion
  • Plan Apo-Chromat 20x/0.8, without immersion
  • LD-C-Apochromat 40x/1.1 w korr UV-Vis_IR, water immersion motorized stage.

Controlled environment:

  • The system is equipped with a large volume incubator from Okolab maintaining temperature at 37°C.
  • Custom design gas micro-chamber is installed. The Gas micro-chamber fits on the microscope stage, inside the temperature controlled enclosure, and hosts the sample. It creates a small volume inside the enclosure in which the pre-mixed and pre-humidified stream of gas is continuously fed.
  • The gas chamber can receive, either two multi well plates or eight 35mm Petri dish or six chambers slides system.
  • The environmental air is controlled with a 3 gas mixer. The mixed gas flows through a humidifier module.

Time lapse videomicroscopes

1 / Nikon

  • The microscope is an inverted Eclipse Ti from Nikon.
  • Imaging software: NIS Elements
  • Camera: ANDOR Technology, Clara Series.
  • Microscope with motorized objective nosepiece.
  • PLAN Fluor 4x/0.1 PhL DL
  • PLAN Fluor 10x/0.3 Ph1 DL
  • S PLAN Fluor ELWD 20x/0.5 DIC N1
  • S PLAN Fluor ELWD 20x/0.5 Ph1 ADM
  • S PLAN Fluor ELWD 40x/0.6 DIC N1
  • S PLAN Fluor ELWD 40x/0.6 Ph2 ADM
  • All objectives are used without immersion.
  • Motorized stage.
  • Several fluorescences :

 

    SEMROCK Filter Set
Bloc Name Part Number Dichroic Exciter Emitter
1 Empty        
2 DAPI DAPI-5060C-000-ZERO 409 - ∞ 352-402 417-177
3 CFP CFP-2432C-000-ZERO 458 - ∞ 426-450 167-499
4 GFP GFP-3035D-000-ZERO 495 - ∞ 457-487 503-538
5 TRITC TRITC-B-000 562 - ∞ 532-554 573-613
6 mCHERRY mCherry-B-000 593 - ∞ 542-582 603-678
- Cy5 Cy5-4040C-000-ZERO 660 - ∞ 608-648 672-712
- CY3 CY3-4040C-000-ZERO 562 - ∞ 491-571 553-633

 

Six filters were installed on the machine and it is possible to install two more filters.

Controlled environment:

  • The system is equipped with a large volume incubator from LIS (Life Imaging Services) maintaining temperature at 37°C.
  • A gas micro-chamber is installed inside the temperature controlled incubator.
  • The gas micro-chamber can receive multiple stage inserts:
    • PetriDish,
    • Multi well plate
    • Chamber Slides system
    • IVF 4 well Dishes
  • The environmental air is controlled with a constant CO2 percentage. The mixed gas flows through a humidifier module.

2 / LEICA

  • The microscope is an inverted DMIRBE from LEICA.
  • Imaging software : Metamorph with the ND acquisition module
  • Camera: Photometrics CoolSnap ES.
  • Microscope with motorized objective nosepiece.
  • C PLAN 10x/0.22 LMC
  • C PLAN 20x/0.3 LMC
  • N-Plan 10x/0.25 PH1
  • N-Plan 20x/0.40 corr PH1
  • N-Plan 40x/0.55 corr PH2
  • All objectives are used without immersion.
  • Motorized stage.

Controlled environment:

  • The system is equipped with a small incubator from PECON maintaining temperature at 37°C and constant CO2 percentage.
  • 2 Heating inserts: 6 Multi well plates or Petri dish (60mm)
  • Temperature and gas controller come from Pecon.