
Microscope: Zeiss Elyra 7 (SIM2 upgraded) Lattice SIM
Wide field based super resolution scope
LOCATION
BRC 214
PRICING
Rice User Fee: $35/hr
Non-Profit Fee: $54.75/hr
For Profit Fee: $175/hr
Rice Training Fee: $70/hr
Non-Profit Training Fee: $109.50/hr
For Profit Training Fee: $350/hr
CONTACTS
Training Contact:
A Budi Utama - budiutama@rice.edu
ADDITIONAL INFORMATION
The Zeiss Elyra 7 is a wide-field based super-resolution system that uses a specialized 3D Lattice illumination pattern for 3D structured-illumination (SIM) images. It can reach up to 120 nm lateral and ~300 nm axial resolution yields up to 2-fold improvement in voxel resolution in the X, Y, and Z dimensions. However, a new SIM2 module has recently been updated into our Elyra 7 that allows even higher resolution post-processing image at ~60 nm lateral and ~200 nm axial resolution. The Lattice SIM illumination allows a better signal to noise ratio, faster, gentler and can reach up to 255 fps during time lapse acquisition significantly faster than regular laser scanning confocal. It also allows a deeper imaging of samples up to around 70 um thickness in comparison to 20 um of sample thickness in classic SIM. SIM is a technology that can be used with standard fluorophores with the same protocol for confocal or regular wide-field fluorescence sample preparations, making it an easily accessible choice for super-resolution results. Our Zeiss Elyra 7 also allows ApoTome imaging using 40X oil objective and lower. A grid based optical sectioning creates highly contrasted images with resolution comparable to SIM imaging and better than traditional laser scanning confocal. It allows to get lateral resolution ~170 nm and axial resolution ~450 nm. However, with the upgraded SIM2 module, apotome imaging can get lateral resolution down to ~110 nm and axial resolution to ~300 nm. ApoTome imaging is also faster than classic SIM and laser scanning microscopes, allowing for time lapse imaging of faster cellular/tissue dynamics. The Zeiss Elyra 7 system is based on Axio Observer 7 (inverted stand) equipped with Incubator XL dark for live-cell imaging with stage-top incubation system. It also has a Motorized Piezo XY scanning stage with Z-Piezo stage insert. Objectives -420340-9901-000 EC Plan-Neofluar 10x/0.30 M27 -420650-9902-000 Plan-Apochromat 20x/0.8 M27 -420762-9900-000 Plan-Apochromat 40x/1.4 Oil DIC (UV) VIS-IR -420782-9900-799 Plan-Apochromat 63x/1.4 Oil DIC M27 -420780-9970-000 Alpha Plan-Apo 63x/1.46 Oil Corr MAG N.A CORRECTIONS IMMERSION W.D. 10X 0.3 EC Plan-Neofluar Air 5.2 mm 20X 0.8 Plan-Apochromat Air 0.55 mm 40X 1.4 Plan-Apochromat Oil 0.13 mm 63X 1.4 Plan-Apochromat Oil 0.10 mm 63X 1.46 Alpha Plan-Apo Oil 0.10 mm Illumination and Detection • Lasers: 405 nm (50 mW), 488 nm (100 mW), 561 nm (100 mW), 642 nm (150 mW) • Two PCO edge sCMOS cameras with 82% QE and Liquid cooling system with 16-bit dynamic range. Imaging software -ZEN (black edition) with Lattice SIM/Apotome modules for image capture not networked. -ZEN Desk for offline storage and analysis workstation. Networked. Both of workstations are connected by hard-wire to allow instant image processing Notes: SIM is only available using the 63X oil and requires closely matching of refractive indices between the immersion oil and the mounting media of the specimen. A series of images are acquired with the illumination pattern in different orientations, giving spatial information that normally would be lost. There are 15 composite images taken to generate a single 2D Classic SIM super-resolution image. The various illumination patterns require 3 rotations and 5 phase shifts for a total of 15 images in order to reconstruct one super-resolution image. In Lattice SIM, The various illumination pattern orientations doesn’t require rotations and a total of 13 or 15 images is required in order to reconstruct a single 2D Lattice SIM super-resolution image. ApoTome Imaging is available using 10x, 20x and 40X objectives. There are 5 composite images to generate a single 2D ApoTome image. ApoTome mode is a faster mode that forgoes the illumination pattern rotations, instead using 5 phase shifts to quickly acquire 2-dimensional image and facilitate optical sectioning. For additional readings about Zeiss Elyra 7 please find these two PDFs below and also included the link for Lattice SIM2 detailed information. Zeiss - Introduction to Lattice-SIM Product information for Elyra 7 https://www.zeiss.com/microscopy/us/products/super-resolution/elyra-7.html