Zeiss Lightsheet
Zeiss LightSheet
The Zeiss LightSheet Z.1 allows fast 3D fluorescence imaging of live or fixed and cleared specimens up to 1 x 1 x 2 cm with reduced phototoxicity and photobleaching.
LightSheet Fluorescence Microscopy (LSFM)
LSFM is based on the Planar Illumination Microscopy (PIM) technique which works by illuminating a specimen with a thin sheet of light that passes through the whole focal plane in the direction of illumination, and gathers the emitted photons in the orthogonal plane. The Zeiss Z.1 microscope illuminates the specimen from both the left and the right side of the specimen and detects the emitted fluorescence orthogonally through a single detection objective. In addition, the thin light sheet is scanned along the focal plane to cover the whole field of view and provide a more homogeneous field of illumination.

The Micro Imaging Facility’s LightSheet Z.1 is coupled with an Environics 4000 gas mixing system to fully control the levels of O2, CO2 and N2 and a tightly regulated thermo chamber, allowing long term live imaging.
System Specifications
- Dual side illumination
- Ultrafast light sheet Pivot scan
4 axis sample positioning system with XYZ and α-stage: motorized - Overview camera with LED illumination for easy specimen positioning
- Fully equipped for live imaging, temperature and O2, CO2 and N2 controlled
- Software: ZEN Black 2014
Detectors
2x 16-bit sCMOS PCO.Edge cameras for simultaneous dual colour acquisition (1920 x 1920 pixels)
Frame rate from 10 to 30 fps at 1000 x 1000 pixels
Illumination objectives
- Zeiss LSFM 5X/0.1 (in combination with 5X and 10X detection objectives
- Zeiss LSFM 10X/0.2 (in combination with 20X objectives)
- Light sheet thickness 2 μm - ~14 μm
Detection Objectives
Lasers
Filters for emission selection
Laser Blocking Filters (LBF)
Filter Module LBF 405/488/561/638
Filter Module LBF 445/515/561
Filter Module LBF 445/515/638
Environics 4000
The Environics 4000 instrument, fully controlled by a standalone software, allows tight regulation of gas concentration, flow and schedule throughout the imaging session.
Environics 4000 Applications
Live imaging
Developmental processes such as cell migration, gene spatio-temporal expression pattern, organ morphogenesis.
Biological dynamic processes such as blood flow, calcium flux.
3D Cell Culture: organotypic and organoid culture, 3D cell culture proliferation
Fixed/cleared imaging
Fast 3D imaging of in toto specimen fixed and fluorescently labelled for volume analysis, atlas preparation. Larger specimen can now also be imaged after clearing, saving hours of slicing, microscopic slide preparation and classic confocal imaging and reconstruction, which are time consuming methods.
Acquisitions post-processing
ZEN Black dual-side imaging fusion
Huygens Multiview fusion and LSFM-dedicated deconvolution
Arivis Vision4D mosaic reconstruction, 3D/4D visualisation, movie recording.
Citation
[Live] Three-dimension images of [specimen] were acquired on a LightSheet Z.1 (Zeiss). The light sheet was focus onto the specimen with a [nX/NA] illumination objective on both the left and the right sides and images were acquired with a [detection objective description], with a z-step of [… μm] and a pixel size of [… nm]. [precise Multiview angle, mosaic’s tile number…]. Dual-side fusion [+ other postprocessing] was run [online/offline] using [Zen Black/Huygens Fuser].
Micro Imaging Facility Support
When you book in time to use the Innovation Centre's Zeiss Lightsheet, you will receive the following support from the Micro Imaging Facility team:
- Specimen preparation and mounting
- X-Clarity based or passive methods specimen clearing
- Full training for fixed and live imaging
- Full training for image processing including Huygens Fuser based deconvolution and Multiview alignment, Zeiss based dual-side imaging alignment, Arivis based mosaic reconstruction, 3D/4D observations.
- Fee for service
Enquiries
Dr Scott Page, BSc, PhD
Head, Micro Imaging Facility
s.page@victorchang.edu.au
Acknowledgement of Country
The Victor Chang Cardiac Research Institute acknowledges Traditional Owners of Country throughout Australia and recognises the continuing connection to lands, waters and communities. We pay our respect to Aboriginal and Torres Strait Islander cultures; and to Elders past and present.
