Instrument Name:

Zeiss Inverted Structured Illumination Microscope (Elyra 7)  Room: X214

Introduction

Structured Illumination Microscopy (SIM) improves resolution by utilizing non-uniform excitation light patterns with sinusoidal intensity variations and powerful image reconstruction techniques. The Elyra 7 uses Lattice SIM technology, which employs lattice patterns (rather than grid lines) to illuminate the sample area. Compared to traditional SIM, Lattice SIM has the advantage of fast imaging speed and high photon utilization, making it suitable for imaging living cells and obtaining high-resolution images of conventional thin samples. Elyra 7 is powered by a new, groundbreaking two-step image reconstruction algorithm called SIM². In the first step, diffraction order combination, noise removal and frequency suppression filtering are applied, producing a digital SIM point spread function (PSF) used in subsequent deconvolution. The SIM² algorithm outperforms traditional single-step methods in resolution, optical cross-section, and stability. Elyra 7 offers a SIM Apotome mode that uses grid lighting and swiftly adjusts fluorescence signals in various focal planes. By capturing three or five images of different grid phases, the software combines them into a single optical section image. Additionally, Burst mode (up to 255 fps) and Leap mode (imaging every three planes to reduce time and photobleaching) further improve speed. Elyra 7 also includes a single-molecule localization module (SMLM), achieving 20–30 nm lateral resolution.

Application

The Elyra 7 inverted microscope is versatile and compatible with a range of scenes, from wide field to super resolution, for both live cells and conventional sections. It supports common dyes and fluorescent proteins and offers the advantages of fast shooting speed and low light damage. The live cell incubation system accommodates 35mm and 60mm petri dishes as well as standard slides; the O2 control module creates a suitable growth environment for anaerobic cells.

Key Technical Specifications

Laser:

405nm(50mW), 488nm(100mW), 561nm(100mW), 642nm(500mW)

Objectives:

10X (Air, NA=0.30, WD 5.20mm) 10X (Air, NA=0.45, WD 2.00mm) 20X (Air, NA=0.8, WD 0.55mm) 25X (Water/Oil/Glycerin/Silicone, NA=0.8, WD 0.57mm) 40X (Water, NA=1.2, WD 0.28mm) 63X (Oil, NA=1.4, WD 0.19mm) 63X (Oil, NA=1.46, WD 0.10mm)

Imaging Modes:

Lattice SIM, SIM Apotome, SMLM and TIRF

Acquisition Speed:

255 frames per second in Burst mode

Filter Parameters:

DAPI: EX 365, BS FT 395, EM BP 445/50; GFP/mRFP/Alexa 633: EX TBP 483+564+642, BS TFT 506+582+659, EM TBP 526+601+688

Detector:

sCMOS high-speed dual camera, physical pixel 2048×2048, pixel size 6.5µm×6.5µm, QE 82%, sampling depth 15bit

Scan Magnification:

1x and 1.6x

Live Cell Incubation System:

Temperature control: room temperature to 60℃, accuracy 0.01℃; CO2 control: 0 to 8%, accuracy 0.01%; O2 control: 0 to 21%, accuracy 0.01%; Humidity: 50%-70% (37℃)

Adapter:

Compatible with 35mm and 60mm petri dishes, standard slides, small eight-well culture tanks, and multi-well plates

Image Processing Software:

Zeiss ZEN (SIM, SIM2, Burst, SMLM algorithms); Arivis Vision4D (visualization and quantification of 3D/4D datasets, volume fusion, channel offset correction, ML-based segmentation and 3D tracking)