Widefield - ZEISS Elyra 7 Lattice SIM² (Irchel)

Widefield, DIC

Widefield (WF) mode (sample illumination with arc lamp), Laser WF mode (sample illumination with laser)

Lattice SIM

Structured Illumination Microscopy, allowing fast and gentle super-resolution imaging (~120 nm in xy and ~300 nm in z) in 3 dimensions.

Lateral resolution (XY): 120 nm, axial resolution (Z): 300 nm (typical experimental FWHM values with objective lens Plan-Apochromat 63× / 1.40 Oil DIC, subresolution beads of 40 nm diameter and excitation at 488 nm)

Lattice SIM² 

SIM² is a novel, image reconstruction algorithm by ZEISS that increases the resolution and sectioning quality of structured illumination microscopy data. Unlike conventional reconstruction algorithms, SIM² is a two-step image reconstruction algorithm. First, order combination, denoising and frequency suppression filtering are performed. All the effects resulting from these digital image manipulations are translated into a digital SIM point spread function (PSF). The subsequent iterative deconvolution uses this very PSF. Similar to advantages of using experimental PSF for deconvolution of hardware-based microscopy data, the SIM² algorithm is superior to conventional one-step image reconstruction methods in terms of resolution, sectioning and robustness. The new SIM² module doubles the conventional SIM resolution and achieves up to ~60 nmlaterally and ~200 nm axially. The Lattice SIM illumination allows for a better signal to noise ratio, more gentle imaging and can reach up to 255 fps during time lapse acquisition, well suited to capture highly dynamic biological processes in live samples. It further allows deeper imaging of samples up to around 70 µm thickness in comparison to only 20 µm of sample thickness in classical SIM. The speed of this imaging modality is well suited to capturing dynamic biological processes in live samples (significantly faster than regular laser scanning confocal).

Lateral resolution (XY): down to 60 nm, axial resolution (Z): down to 200 nm (typical experimental FWHM values with objective lens Plan-Apochromat 63× / 1.40 Oil DIC, subresolution beads of 40 nm diameter and excitation at 488 nm; Resolution is sample and SNR dependent.)

Apotome

Grid-based optical sectioning to create highly contrasted images with high lateral and axial resolution. The Apotome mode is also amenable to SIM2 processing, allowing for high speed acquisition with high contrast, high resolution and low phototoxicity. In combination with 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.

Lateral resolution (XY) of 140 nm, axial resolution (Z) of 275 nm for 40×; lateral resolution (XY) of 285 nm, axial resolution (Z) of 550 nm for 20×; lateral resolution (XY) of 710 nm, axial resolution (Z) of 1300 nm for 10×
 

SMLM

Single-molecule localization microscopy like dSTORM, PALM and PAINT allowing resolution down to ~20-30 nm in xy and ~50-80 nm in z.  

HILO & TIRF

Highly inclined and laminated optical sheet (HILO) and total interbal reflection illumination (TIRF).