Marianas LightSheet

Versatile Multiview Light Sheet Microscopy System for Imaging Model Organisms

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Versatile Multiview Light Sheet Micrsocopy System for Imaging Model Organisms

Marianas LightSheet (MLS) combines the low phototoxicity and large sample space of dual inverted selective plane illumination microscopy (diSPIM) with the power and flexibility of a research-grade inverted microscope system. MLS supports a wide variety of specimen types and sizes as well as experimental designs including photoablation, photomanipulation, and computer-generated holography. Add spinning disk confocal, TIRF and FLIM to transform the system into a versatile multimodal imaging system all controlled with SlideBook software.

Slide

Environmental Control

Stage top and cage incubators
Temperature, gas and humidity

Light Sheet Scanners

Objective Piezos

Motorized SPIM Z Drive

High-speed MEMs mirror or cylindrical lens scanners

95% QE low read noise
Large field-of-view options

150µm or 300µm piezos for long-term high-speed 3D sampling

Water and multi-immersion dual-view optimized objective pairs

Motorized objective, condenser and path selection
Autofocus (Definite Focus 3)
PSF-optimized objectives

High precision multi-position and vertical montage capture

Scan-Optimized X,Y Stage

LaserStack

Fiber Switcher

Additional Light Path

Millisecond timing and trigger
Control of multiple devices

Modular laser combiner
Up to eight lasers

Up to four fiber outputs
Millisecond path switching

Add cameras, photomanipulation devices,
spinning disk confocal and more

High-precision movement of sample during long-term experiments

Live cell 3D confocal imaging
Super-resolution dual microlens disk

Ablate

Laser ablation system
355nm and 532nm

sCMOS Cameras

Vector2

Vector3

Objectives

Fully Automated
Research Microscope

Epi mounted or camera port
mounted photomanipulation
Modular high-speed X,Y scanner

Epi-mounted TIRF & photomanipulation
Spinning TIRF with expansive FN22 FOV
Liquid light guide input for LED light source

Highlights

Scanning Modes

Fixed

Fixed

Ultra-fast single plane time-lapse capture with stationary light sheet.
ZEBRAFISH HEART
Zebrafish heartbeat (GFP-labeled Cardiomyocytes). Courtesy of Dr. Jamie Nichols, University of Colorado.
 
Piezo

Piezo

Fast 3D capture up to 3 volumes/sec of smaller specimens with synchronized sheet and objective movement.
ARABIDOPSIS ROOT
Piezo slice scan capture of an Arabidopsis root (YFP-labeled membrane). Courtesy of Dr. David Ehrhardt, Carnegie Institution for Science.
 
Stage

Stage

Large 3D capture up to 1 volume/sec of specimens translated through a stationary light sheet.
CLEARED BRAIN SLICE
Stage scan captures of a cleared mouse brain (GCaMP-GFP) with a pair of 10x (top) and 40x (bottom) objectives. Courtesy of Dr. Rob Campbell, University College London.
 
Dual-View Piezo

Dual-View Piezo

Alternating Piezo captures combined with SlideBook Multiview Reconstruction for isotropic resolution.
DICTYOSTELIUM
Dual-view piezo slice scan timelapse of Dictyostelium cells (membrane-rhodamin).
 
Dual-View Stage

Dual-View Stage

Alternating Stage captures combined with SlideBook Multiview Reconstruction for isotropic resolution.
ZEBRAFISH
Dual-view stage scan timelapse (GFP-actin). Courtesy of Elric Esposito, Spencer Shorte UtechS Photonic BioImaging and Nicolas Dray, Laure Bally-Cuif Zebrafish Neurogenetics lab, Institut Pasteur.
 
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Fixed
Fixed
Piezo
Piezo
Stage
Stage
Dual-View Piezo
Dual-View Piezo
Dual-View Stage
Dual-View Stage

Objective Pairs

Magnification Numerical Aperture Refractive Index Maximum Field of View Recommended Use
Water Immersion
10x 0.3 1.33-1.4 1490µm Largest field of view
20x 0.5 1.33-1.4 620µm Balanced field of view and resolution
40x 0.8 1.33-1.4 370µm Highest Resolution
Multi-Immersion
10x 0.3 1.33-1.56 1470µm Largest field of view
16x 0.4 1.33-1.56 850µm Higher resolution large field
24x 0.7 1.33-1.56 590µm Highest resolution

Flexible Sample Mounting

Multimodal Platform

Combine Marianas LightSheet with spinning disk confocal, holography and ablation systems for incredible flexibility on one system. Seamlessly control and switch imaging modes via SlideBook. LaserStack with FiberSwitcher enables millisecond switching of up to 7 lasers across modalities.

Technology

Dual Inverted Selective Plane Microscopy

Dual inverted selective plane microscopy (diSPIM) uses dual camera and scanner objective pairs positioned at 45° to the specimen plane to alternate capture and excitation between Path 1 and Path 2. The flexible system geometry allows for single- and dual-sided imaging with conventional sample mounting. SlideBook seamlessly controls all hardware modalities, deskewing, stitching/montaging, Multiview Reconstruction and joint deconvolution resulting in isotropic sub-cellular resolution across a wide range of samples. Marianas LightSheet integrates diSPIM with a research-grade inverted microscope enabling complex multi-modal light sheet experiments with photoablation, photostimulation and computer-generated holography.

ZEBRAFISH
Dual-view stage scan capture of a Zebrafish (GFP-Actin). Path 1 (left) and path 2 (right). Courtesy of Dr. Cody Smith, University of Notre Dame.

SlideBook Multiview Reconstruction

SlideBook for MLS supports several methods to deskew, rotate, register and deconvolve diSPIM images. Dual-view capture registration is performed either through Point of Interest Detection or 3i’s proprietary Local Cross-Correlation and Registration algorithms. Results are instantly displayed in 3D for quick adjustment to achieve an ideal fit before joint deconvolution produces a fused image. Large datasets that would otherwise require high-powered computer resources can be automatically split into a computer’s available RAM and computed in parallel utilizing GPU-optimized processes.

SlideBook Software for Acquisition and Analysis

SlideBook software supports research microscopy through the entire experimental process. By managing everything from instrument control to image processing and data analysis, SlideBook allows scientists to focus on investigation rather than instrumentation. SlideBook controls hundreds of instruments in and around the microscope from dozens of manufacturers enabling researchers to integrate their preferred components and upgrade to the latest devices once available.

Slide

User-Selectable App Appearance

Select a color scheme from dozens of options
Switch on-the-fly from dark to light themes

SlideBook Open File Format

Directory-based open file format for big data and high performance computing applications

Multiview Reconstruction

SlideBook can use a number of methods to deskew, rotate, register and deconvolve images captured with MLS. Data is visualized, deskewed and rotated with a single click. Initial shifts and registration results can be instantly checked in 3D and adjusted before joint deconvolution to a fused image.

NVIDIA CUDA GPU Acceleration

GPU acceleration of computationally-intensive operations such as deconvolution

Volume Rendering

3D and 4D visualization tools rotate and deskew MLS data on-the-fly and support a user specified bounding box and storyboard interface where multiple perspectives can be assembled into a single movie.

3D Capture Status

Multi-Position and Montage

Volumetric projection during 4D capture supported across all instruments

Capture single timepoints or time-lapses at multiple positions with different experimental setups and throughout the range of capture modes

Capabilities

Capture

Control hundreds of devices including microscopes, stages, lasers, wheels, piezos, scanners, shutters and much more.

View

Visualize data through any numbers of portals, from single images to z-stacks, time lapse, color channels and 4D views.

Analyze

Analyze images and extract statistical data via a wide variety of algorithms while maintaining original data integrity.

Scripting

Macro scripting for capture and analysis enhances the flexibility and power available to users.

Communicate

Present and export data easily as 16-bit TIFFs, 3D movies, graphs or spreadsheets. Data is directly portable to MATLAB and Excel and adheres to Open Microscopy Environment (OME) standards.

Partners

MATLAB

Through hierarchical and conditional capture, user-supplied MATLAB programs can control experimental workflows.

Microvolution

Microvolution software delivers nearly instantaneous deconvolution by combining intelligent software programming with the power of a GPU.

Aivia

Aivia is an innovative and complete 2D-to-5D image visualization, analysis and interpretation platform with artificial intelligence-guided image analysis.

Dell

The latest high-power computer workstations control all microscope hardware and enable high-speed processing, segmentation and volume rendering of terabyte (TB) datasets.

Powerful Computer Workstation

3i provides high-power computer workstations to control all microscope hardware necessary for acquisition as well as enable processing, segmentation and volume rendering of terabyte (TB) datasets without additional computer resources.  Solid state drives in RAID configurations provide high-speed storage for capture, while traditional high-capacity hard drives provide longer-term storage of datasets. NVIDIA Quadro GPUs work in parallel with the latest Intel Xeon processors for fast stitching, processing and rendering of captured data.

Petabyte Data Storage

3i offers DDN® unified storage systems to allow direct acquisition and analysis, without time-consuming file transfers, at volumes ranging from 500TB to over 2PB. DDN storage systems are an ideal choice for labs and facilities looking to optimize acquisition workflows and/or incorporate data analysis pipelines.

Data Sets

DROSOPHILA EYE
Stage scan Y-montage of a Drosophila pupal retina showing utrophin labeled F-actin in pseudocolour. Courtesy of Courtney Lancaster, Pichaud lab, University College London Laboratory for Molecular Cell Biology (LMCB).
HUMAN EMBRYONIC KIDNEY (HEK) CELLS
Dual-view piezo scan capture of HEK cells with labeled tubulin (green), actin (magenta) and nuclei (blue). Courtesy of Dr. Deirdre Kavanagh, University of Birmingham.
 
biofilm
Dual-view stage scan of Candida albicans growth (GFP-Mitochondria). Courtesy of Dr. D Pentland, collaboration Gourley-& Laissue-Labs, Universities of Kent & Essex.
 
zebrafish tail
Dual-view stage scan capture of a Zebrafish (GFP-Actin). Courtesy of Dr. Cody Smith, University of Notre Dame.
 
cyanobacteria
Maximum intensity projection (MIP) of a stage scan timelapse capture of cyanobacteria biofilm autofluorescence excited with 488nm and 561nm after 48 hours. Courtesy of the Institute of Cancer Research, London.
 
drosophila
Dual-view stage scan capture of a Drosophila wing disc showing Viking (magenta) and membrane (green). Courtesy of Dr. Yanlan Mao and Dr. Rob Tedley, University College London Laboratory for Molecular Cell Biology (LMCB).
ARABIDOPSIS EMBRYO
Dual-view piezo scan capture of an Arabidopsis embryo showing autofluorescence excited with 405nm (blue) and 488nm (green). Courtesy of University of Warwick.
zebrafish embryo
Stage scan capture of a zebrafish embryo 20 hours post-fertilization with labeled actin, tubulin and nuclei.
 
drosophila embryo
Piezo scan timelapse of synchronised cell division of Drosophila melanogaster embryo (~ 4hours old, H2B- labeled chromatin).
 
arabidopsis trichomes
Piezo scan timelapse of Arabidopsis trichomes (GFP labeled actin).
 
mouse brain
Stage scan capture of iDISCO-cleared embryonic mouse brain with antibody stainings for smooth muscle cells (white) and endothelial cells (magenta). Courtesy of Tijana Perovice, Gerhardt lab, Max-Delbrück Center for Molecular Medicine.
 
drosophila embryo
Stage scan capture of Drosophila embryo with labeled nuclei (orange) and corpora allata cells (blue). Courtesy of Dr. Lucas Dent, the
Institute of Cancer Research, London.

Additional Resources

Brochures

Marianas LightSheet

Marianas SDC

Webinars

 

Marianas LightSheet: Selective Plane Illumination with Isotropic Resolution

Find out more about Marianas LightSheet









    SALES

    Email: sales@intelligent-imaging.com

    Phone: +1 (303)-607-9429 x1

    SUPPORT

    Email: support@intelligent-imaging.com

    Phone: +1 (303)-607-9429 x2