The Light Microscopy Facility offers one-on-one technical support, consultation, and training to ensure optimal use of our many advanced capabilities. Please, contact us to help you identify the best technology for your imaging needs.
Airyscan technology: Super Resolution and High Sensitivity
The Zeiss AiryScan technology uses multipoint confocal acquisition combined with deconvolution and pixel reassignment to achieve isotropic gain of resolution with a factor 1.4x in 2D acquisitions and 1.7x in 3D acquisitions. Gain of resolution can be achieved routinely without requiring any specific sample preparation protocol. Super resolution can be maintained even ~50+ microns into a scattering sample in the presence of substantial out-of-focal-plane fluorescence. The Airyscan technology improves significantly the detection sensitivity compared to conventional detectors. Higher SNR can be achieved with gentler excitation which makes it a technology of choice for dim samples and live experiments.
Confocal microscopes, wide-field Apotome microscope, and multi-photon microscope each use different technologies to acquire optical sections that can be combined into a 3-dimensional image. We support several software packages that offer 3D visualization capabilities.
3D imaging capabilities will be expended very soon with custom Lattice Light Sheet Microscopes and Zeiss Light Sheet Z1.
Light Sheet Microscopy
Light Sheet microscopy allows fast 3D imaging of large volumes with gentle excitation.
- The Zeiss Light Sheet Z1 will provide a turn-key instrument for routine high-speed imaging with intermediate Z resolution.
- The Lattice Light Sheet Microscope (LLSM) will provide a custom system for high-resolution 3D imaging. This instrument has been designed to accommodate large samples for Expansion Microscopy.
These systems are particularly useful on cleared samples, expended samples, transparent organisms but also on cell culture for the LLSM.
All of our microscopes can automatically capture series of images over time. Frame rates depend on the technology used and signal strength. Wide-field images with triggered acquisition can exceed 50 fps, while confocal imaging can rarely exceed 2 fps. Higher (100s fps) frame rates are possible if imaging is restricted to only a smaller number of pixels. Only some microscopes are equipped with temperature and CO2 controlled environment.
Spectral imaging consists in the measurement of the fluorescence emission spectra at each pixel in an image. Using reference emission spectra, linear unmixing allows separating the contribution to intensity of spectrally overlapping dyes. Linear unmixing can be used to image dyes with overlapping emission spectra, to image samples with more than 4 dyes, to speed up acquisitions by acquiring all the dyes in parallel and unmix them post-acquisition, to differentiate specific dyes emission from auto-fluorescence.
High Throughput Imaging
Two slide scanners are available with up to 200-250 slides in a single run. The TissueGnostics slide scanner can acquire images in regular wide-field mode or in confocal spinning disk mode. The spinning disk optical sectioning increases contrast by rejection of out-of-focus light.
High Content Imaging / Automated acquisitions
Several of our systems are equipped to design automated acquisitions of custom sample carriers or multi-well plates. Complex acquisition protocols can be carried out without requiring any coding. High Content Analysis (HCA) software with heat maps representation is also available to quantify automatically multi-plate acquisitions.
Two fluorescence stereoscopes offer binocular disparity-based visualization of large (>100 um) samples at lower magnification. Both low and high power objectives are available.
Custom Image Processing and Analysis
We can provide support for image processing and quantification. Customized macros or plugins can be written for ImageJ. These processes can be fully automated if necessary. Please contact us in advance, as suitable image acquisition settings are a prerequisite for accurate image analysis.
We fully support several analysis software packages including ImageJ, FIJI, Nikon Elements, and TissueGnostics.