Azimuthal TIRF iLas2 module on a Zeiss Axio Observer 7 microscope

Applications

This microscope allows three-color TIRF imaging on living samples, in physiological conditions. The iLas2 module from Gataca Systemps consists in an azimuthal TIRF: the rotating laser allows homogeneous imaging of the sample and elimination of interference fringes. TIRF angle can be controlled independently for each laser. Z-stacks can also be obtained, allowing 3D reconstruction on 800 nm depth with an axial resolution of 50 nm. Importantly, the microscope is equipped with Definite Focus 3 for drift correction during time-lapse acquisitions. Of note, the microscope can also be used in a classical wide-field mode, allowing up to 3-color imaging of fixed and living samples in DAPI, GFP and Cy3.

Azimuthal TIRF iLas2 module on a Zeiss Axio Observer 7 microscope

Technical description

Inverted Zeiss Axio Observer 7 stand, with fully motorized stage (XYZ)
Equipped with azimuthal TIRF module (iLas2) from Gataca Systems
Lasers (diodes): 488 nm, 561 nm, 639nm
Camera: sCMOS Photometrics Prime BSI Express (QE 95%, pixel size 6.5µm x 6.5µm)
Objectives: 20x (EC Plan-Neofluor, NA 0.5, dry), 100x (Alpha Plan-Apochromat, NA 1.46, oil)
Drift correction: Definite Focus 3
UV-Vis light source: HXP 120V (Hg lamp)
Filters: single-band filter cubes for DAPI, GFP and DsRed, quadband filter cube, wheel with emission filters (single-band filters for DAPI, GFP, Cy3 and Cy5 + quadband filter).
Camera: sCMOS Photometrics Prime 95B (QE 95%, pixel size 11µm x 11µm)
Equipped with a stage top incubator for temperature/CO2/humidity control (Tokai Hit)
Antivibration table (compressed air)