Advanced Medical BioImaging Core Facility 

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ZEISS Lattice Lightsheet 7

Charité Campus Mitte, CCO Rm. 01-307, Virchowweg 6, 10117 Berlin, Germany

Dear All,

we are happy to announce that the Zeiss lattice light sheet (LLS 7) has been installed in the new AMBIO facility lab room 01-307 located next door in the CCO building. 

The main advantage of this system over fast live-cell imaging systems (like the spinning disk confocal - SDC) is that the photodamage is minimized due to the light sheet illumination (s. schematic below). Therefore, long-term ultra-fast 3D live-cell imaging at subcellular resolution without remarkable phototoxicity and bleaching is now possible. The SDC has slightly better resolution but more photo damage compared to the LLS. So, if the SDC is damaging your sample, the LLS might be the solution for you. 

The LLS allows standard imaging sample carriers (slides, 35 mm cell culture imaging dishes, well plates). The system provides near-isotropic resolution along X/Y/Z-axes. Available imaging lasers are 488 nm, 561 nm, and 640 nm. Additionally to this basic LLS system, it is equipped with a laser scanning-based photo activation/manipulation system with the following activation/ablation lasers:

  • 1 photon CW excitation at 405 nm or 477 nm (bleaches/ablates a cylindrical hole into the probe)
  • 2 photon pulsed excitation at 780 nm (for activation/ablation in a 3D focal spot!)

Further information about the LLS7 can be found below and at the product homepage.

Comparison of current 3D imaging systems at AMBIO

Applications and Samples

Specifications

Imaging Modes

  • Transmitted light with oblique contrast for sample positioning and overview
  • Reflected light and epifluorescence with laser for beam adjustment and fluorescence light sheet imaging
  • ibidi stage top incubation system (temperature, CO2, humidity) and integrated auto-immersion mechanism enable unattended long-term experiments
  • Photo-manipulation: 405 and 473 nm and lasers for FRAP and photo-activation/photo-switch and 780 nm laser for ablation experiments

Acquisition

  • Speed: Volume: 3 Vol/s @300 μm x 50 μm x 20 μm, plane: 400 frames/s @300 μm x 20 μm
  • Penetration depth up to 80 - 200 μm

Illumination

  • LED (white & red) for transmitted light
  • Laser for reflected light & epi-fluorescence: 488 nm (2 mW), 561 nm (2 mW), 640 nm (1 mW)

Detectors

  • pco.edge 4.2 CLHS sCMOS camera (2048x2048 pixels, 300 x 300 µm, 100 fps@16-bit full frame, 82% QE, 6.5 µm pixel size)

Software

  • ZEN 3.5 (blue edition)
  • Lattice Lightsheet Processing Module (deskewing, coverglass transformation)
  • arivis Vision4D® software for advanced stitching, channel shift, high resolution volume rendering, and powerful deconvolution

Special

  • Ultra-fast live 3D subcellular imaging with minimal phototoxicity and bleaching
  • Near-isotropic resolution along the X, Y, and Z axes
  • Using standard coverslips, 35 mm cell culture imaging dishes or well plates
  • 5-axes stage tilt-able with high precision in X and Y

 

Objectives/Optics

  • Illumination (light sheet): 13.3x / NA 0.4
  • Detection: 44.83x / NA 1.0