Microscope: Zeiss Elyra 7 (SIM2 upgraded) Lattice/SIM with TIRF
Wide field Lattice/SIM based super-resolution with TIRF microscope
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 the Elyra 7 that allows even higher resolution post-processing at ~60 nm lateral and ~200 nm axial resolution. 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) -ZEN Desk for offline storage and analysis workstation. Networked. Both of workstations are connected by hard-wire to allow instant image processing Notes: 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. TIRF microscopy can be performed only with Alpha Plan-Apo 63x/1.46 NA Oil Corr objective. 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
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.
Lattice/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 rsoltuion 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 microscope also allows you to work with Total Internal Reflection Fluorescence (TIRF) microscopy technique using the specifically approved high NA Alpha Plan-Apo 63x/1.46 NA Oil Corr. TIRF microscopy is commonly used to observe a thin region of a specimen with usually less than 200 nm image section.
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 (for Lattice/SIM)
-420780-9970-000 Alpha Plan-Apo 63x/1.46 Oil Corr (for TIRF only)
• 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.
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.