Light Sheet · Zeiss
A mesoscale light sheet fluorescence microscope designed for fast, gentle 3D imaging of large cleared or intact specimens. Ideal for whole-organ, whole-embryo, and large tissue imaging with minimal phototoxicity.
Instrument Features
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Mesoscale Sample Chambers
Large sample chambers accommodate whole cleared organs (brain, kidney, embryos), intact zebrafish, or large tissue blocks. Supports CLARITY, iDISCO+, CUBIC, and other clearing method samples in appropriate immersion media.
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Dual-Sided Illumination
Two opposing light sheet arms illuminate the sample from both sides simultaneously. Dual-side illumination eliminates shadowing artifacts common in single-sided LSFM and provides uniform signal across large specimens.
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sCMOS Camera Detection
High-sensitivity scientific CMOS cameras provide fast, low-noise detection. The entire focal plane is captured simultaneously (no scanning), enabling volume acquisition rates orders of magnitude faster than point-scanning confocal.
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Gentle Imaging — Low Phototoxicity
Sheet illumination confines light to only the detection plane, dramatically reducing total sample exposure compared to confocal. Essential for live imaging of embryos and developmental specimens, and for preserving signal in cleared tissue.
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Multiple Objective Options
Detection objectives range from 5× to 20× to accommodate specimens from mm to cm scale. Matched illumination objectives shape the light sheet to the detection field of view for optimal resolution throughout the sample.
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Multi-Channel 3D Acquisition
Simultaneous or sequential multi-channel Z-stack acquisition of entire specimens in minutes. Tiling support allows coverage of specimens larger than a single field of view. ZEN software enables automated multi-position and time-lapse experiments.
Startup Guide
Power On System
Turn on the main system, laser launch, and camera. Launch ZEN Black (Lightsheet configuration). Allow 15–20 min for initialization.
Prepare the Sample Chamber
Select the appropriate chamber for your sample size and clearing method. Fill with the correct immersion medium (e.g., BABB for iDISCO, 80% glycerol for CUBIC, water for aqueous samples). Ensure the medium RI matches your detection objective specification.
Mount Your Sample
Attach your cleared tissue or specimen to the sample holder using low-melting agarose, mounting putty, or a custom holder. Ensure the specimen is stable and will not drift during acquisition. Submerge into the chamber.
Select Objectives
Choose the appropriate detection objective (e.g., 5× Clarity, 20× Plan) and matching illumination objectives. Enter the objective and immersion medium parameters in ZEN to set the correct working distances.
Activate Lasers & Configure Channels
Enable the required laser lines. Set detection filter cubes for each channel. Configure single or dual-side illumination based on specimen size and transparency.
Navigate & Find Focus
Use Live mode to navigate to your specimen. Adjust the Z position and sample orientation (using rotational mount if available). Align the light sheet to the detection focal plane using the pivot scan and sheet thickness controls.
Set Z-Stack Parameters
Define the top and bottom Z limits of your acquisition. Set Z-step size (typically 2–5 µm for mesoscale imaging). Enable dual-side illumination fusion for uniform coverage. Set exposure time per channel.
Acquire
Start the Z-stack acquisition. Monitor the first few slices for focus and signal quality. Large specimens may generate files of 10–100+ GB — ensure sufficient storage is allocated before starting.
Shut Down
Remove sample from chamber. Rinse chamber with appropriate solvent if organic clearing agents were used (BABB/DBE are hazardous — follow posted chemical safety procedures). Turn off lasers and system per shutdown checklist.
⚠️ Cleared tissue samples using organic solvents (iDISCO/BABB) require chemical-resistant materials and specific safety protocols. Contact Adeela before your first cleared tissue session.