- LSE Center Publications
- Microscopy & imaging
- Super Resolution Microscope: Lattice SIM/STED/STORM
- Zeiss- Light Sheet Z7
- Nikon- Spinning Disk Confocal
- LSM 880- Upright confocal with MP laser
- LSM 710 – Inverted confocal
- LSM 700 – Inverted confocal
- LSM 980- Inverted confocal with Airyscan2
- Cytation™ 5 – BioTek
- Leica DMI8 Inverted Fluorescent Microscope
- Olympus Fluorescent Binocular Microscope
- Image analysis & processing
- Flow Cytometry
- Virus room – Emerson Building
Super Resolution Microscope: Lattice SIM/STED/STORM
Description
Multi Modality Super Resolution (SR) microscope that combined lattice SIM, 2D-STED and STOTM/PALM methods for SR, this system is designed for live-cell imaging.
The Elyra 7 eLS is flexible platform featuring unique lattice structured illumination (SIM^2) for fast and gentle 3D super-resolution on live samples as well as Single Molecule Super-resolution imaging. Lattice SIM^2 provides optical sectioning and a doubling of diffraction-limited resolution in 3D (60 nm in xy and 150 nm in z). SMLM part allows for resolutions in the typical range of 20-30 nm laterally and 50-80 nm axially. The system features groundbreaking light efficiency and allows for gentle live cell imaging at speeds of dozens of frames per second over time. It includes state-of-the-art sCMOS cameras for large field of view imaging, SIM- and SMLM optimized optics, focus drift compensation control and
full incubation.
The STEDYCON is a confocal microscope and STED nanoscope with a resolution down to 30 nm. It have a super-intuitive user interface by that the STEDYCON provides an intelligent microscope platform.
Features
SYSTEM SPECIFICATIONS:
Super Resolution (SR) imaging: Lattice SIM^2 (Zeiss), STORM (Zeiss) and STED (Abberior)
Confocal imaging : StedyCon (Abberior- confocal)
Z Stack imaging: Apotom (Zeiss)
TIRF imaging:
Stand: Zeiss Axio Observer 7 SR RP stand for ELYRA 7
XY-stage: motorized
Z – Z PIEZO WSB 500 um range
Definite Focus 2 (Zeiss)
StedyFocus (Abberior)
HXP fluorescence illumination
Equipped for live imaging (temperature and CO2 incubation)
Software: ZEN
LASER OPTIONS:
The Zeiss system includes 4 visible solid state lasers:
– UV diode laser – 405 nm (50mW)
– Blue diode laser 488 (500mW)
– Green diode laser – 561 nm (500mW)
– Red diode laser – 642 nm (500mW)
The Abberrior system includes 4 visible solid state lasers:
– UV diode laser – 405 nm (continues)
– Blue diode laser 488 (pulse)
– Green diode laser – 561 nm (pulse)
– Red diode laser – 640 nm (pulse)
STED laser depletion: 775nm (pulsed)
CAMERA SPECIFICATIONS:
Model: Two pco.edge sCMOS (version 4.2 CL HS)
Effective number of pixels: 2048 x 2048
Pixel size: 6.5µm x 6.5µm
Effective Area: 13.312mm x 13.312mm
Exposure times from 100 µs to 20 s
Dynamic range: 16 bit
Max frame rate: 100 fps @ full frame (CL/CLHS)
40 fps @ full frame (USB)
Quantum efficiency (peak): > 82% (@590nm)
Read out noise: extreme low readout noise of 0.8 e.
Catalog # | 420340-9901-000 | 420650-9902-000 | 420762-9900-000 | 421787-9970-799 | 420782-9900-720 | 420780-9971-000 | 420792-9800-720 |
Magnitude | 10x | 20x | 40x | 63x | 63x | 63x | 100x |
Type | Plan-Apochromat | Plan-Apochromat | Plan-Apochromat | C-Apochromat | Plan-Apochromat | alpha Plan-Apochromat | alpha Plan-Apochromat |
NA | 0.3 | 0.8 | 1.4 | 1.2 | 1.4 | 1.46 | 1.46 |
Immersion | Dry DIC |
Dry DIC |
Oil DIC |
Water DIC |
Oil DIC |
Oil DIC |
Oil DIC |
Imaging Application | WF/FL | WF/FL/ Apotome |
WF/FL/ Apotome |
WF/FL/ Apotome/ Lattice SIM |
WF/FL/ Apotome/ Lattice SIM |
WF/FL/ Apotome/ Lattice SIM/ STED/TIRF |
WF/FL /Apotome/ STORM/PALM/ STED/TIRF |
CAMERA FILTERS :
Beam Splitter | LP 560 | BP490-560/ LP640 | ||
CAMERA | CAM1 | CAM2 | CAM1 | CAM2 |
Detection | BP 570-620 | BP 420-480 | BP 495-550 | BP 420-480 |
Detection | LP 655 | BP 495-550 | LP 655 | BP 570-630 |
Detection | — | — | — | LP 740 |
Acquisition & Analysis:
- Acquisition is performed using the ZEN black software which provides wide range of image processing functions: 2D/3D, projection, reconstructing, co localization, intensity measurements and more. The flexible secondary dichroic beam splitter makes this system fully capable of acquiring super resolution and performing dual camera imaging. in addition, Elyra 7 with Lattice SIM^2 takes you beyond the diffraction limit of conventional microscopy to image your samples with superresolution. Elyra 7 is the fastest processes in living samples – in large fields of view, in 3D, over long time periods, and with multiple colors. The new Lattice SIM technology of Elyra 7 brings structured illumination microscopy (SIM) to a new level. Groundbreaking light efficiency gives you gentle superresolution imaging with incredibly high speed – at 255 fps with imaging speed faster than ever before.Elyra 7 combine Lattice SIM^2 with single molecule localization microscopy (SMLM) for techniques such as PALM, dSTORM and PAINT. Enabled to choose among your labels when imaging with resolutions down to 20 nm laterally. High power laser lines allow to image sample, from green to far red.In addition, it enabled a wealth of contrasting techniques and combine them with optical sectioning. The new Apotome mode gives superfast optical sectioning of your 3D samples.
- The STEDYCON software: only minutes of training are required to acquire STED and confocal images. Thanks to the powerful dye database behind, only minimal user input is needed, and even with unkown samples, you get with 3 clicks to the first STED image!
Applications
The system enables amongst many others, the following applications:
- Super-Resolution Imaging:
Lattice Structure Illumination Microscopy (Lattice SIM^2)
STimulated Emission Depletion (STED) Microscopy :2D STED
Single Molecule Localization Microscopy (SMLM): PALM/dSTORM/PAINT - Section Imaging (Z stack):
Apotome -Zeiss
Confocal -Abberior - Total Internal Reflection Fluorescence (TIRF) Microscopy
- Fast Image Acquisition for Live Cell Imaging and Time-Lapse studies
- Z-Stacks for localization of cells in living tissues and 3-D reconstruction
- Super-resolution localization of sub-cellular compartments and quantities co-localization
- FRET – fluorescence resonance energy transfer
- FRAP – fluorescence recovery after photobleacing in a super-resolution microscopy
- PhotoActivation and PhotoManipulation in a super-resolution microscopy
* Mark and find feature – The motorized stage allows marking specific coordinates using small magnifications or tiling of large scanning areas for further detailed scanning.