Equipment

AMCF IMAGING TECHNOLOGIES

SUPER-RESOLUTION TO MICROSCOPIC TO MESOSCOPIC IMAGING


*Adapted from Miltenyi Biotec

ZEISS ELYRA PS.1 SUPER RESOLUTION MICROSCOPE

 

The Zeiss ELYRA PS.1 is an inverted microscope designed for super-resolution (SR) structured illumination microscopy (SIM) and single-molecule localization microscopy (SMLM) inlcuding PhotoActivated Localization Microscopy (PALM) using photo-switchable/convertible fluorescent proteins, Total Internal Reflection Fluorescence (TIRF) and STochastic Optical Reconstruction Microscopy (STORM) using specific fluorescent organic dyes. The Elyra S.1 can image four fluorescent colors, roughly corresponding to DAPI, Alexa488/GFP, RFP/Alexa568, and CY5/A647 (see below) during SIM imaging, PALM/STORM imaging can capture 2 channels. SIM uses structured light to excite and collect (traditional fluorophores) in small areas that are rotated and accumulated overtime to generate a composite SR image. PALM/STORM requires specific photo-activated/photo-converted/blinking fluorophores (see AMCF Imaging Resources) that are slowly accumulated to generate a composite SR image. Due to intense light and longer collection times during SR imaging, rapidly bleaching and/or dim fluorophores will not work.  Contact the AMCF to learn more and/or schedule your reservation.

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ZEISS LSM 800 WITH AIRYSCAN FOR HIGH RESOLUTION IMAGING

The Zeiss 800 CLSM with Airyscan is an inverted microscope with transmitted (HAL), UV (HBO) and laser excitation for collection transmitted light images (bright field and DIC) and/or confocal fluorescence images. This system dramatically increases conventional confocal image resolution to ~180 nm using Airyscan technology distributing light across an array of detectors (see link in system highlights). Fast linear scanning and high-sensitivity GaAsP detectors increase the imaging speed and sensitivity for most imaging applications including, multi-channel, co-localization, live cell, 3D, and time series imaging. Enhanced sensitivity supports use of lower laser powers to reduce photobleaching and phototoxicity during live cell imaging. Contact the AMCF to learn more and/or schedule your reservation. Reference figure, Korobchevskaya et al. 2017.

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ZEISS 710 CONFOCAL LASER SCANNING MICROSCOPE

The Zeiss 710 LSM is an inverted microscope with transmitted (HAL), UV (HBO) and laser excitation to collect transmitted light images (bright field and DIC) and/or confocal fluorescence images. This system supports most basic imaging applications, multi-channel and spectral, co-localization, live cell, 3D, and time series imaging. Contact the AMCF to learn more and/or schedule your reservation. Fluorescent image, Muthuraj et al. 2021.

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ZEISS CELLDISCOVERER 7

 

The Zeiss Celldiscoverer 7 is a widefield imaging system for automated, time-lapse imaging of live samples. Samples may be imaged on slides, dishes, and across an array of multi-well dishes including 96 well plates. The system offers a temperature and C02 controlled incubation system for optimal short- and long-term live sample imaging. ZEN Blue allows users to calibrate the system to their sample carrier, input exact imaging parameters, and create a focus strategy keeping the sample in focus across locations and timepoints. While widefield imaging offers accelerated imaging of larger views of view at lower resolutions when compared to confocal microscopy, this system is equipped with afocal magnification changers for high resolution imaging of smaller fields of view that can be subsequently stitched together to generate larger field of view composite images. Contact the AMCF to learn more and/or schedule your reservation.  

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ZEISS AXIOSCAN 7 WHOLE SLIDE IMAGING SYSTEM

Coming Soon: Installation Spring/Summer 2022!

The Zeiss Axioscan 7 is a high-performance whole slide scanning system for fluorescence, brightfield, and polarization imaging. Up to 100 slides can be imaged using customized scanning profiles using a specalized slide cassette system which means slides remain upright and untouched during the imagign process. Fluorescence and brightfield imaging modes can be used idependently or combined to maximize data collected from your samples. The new transfer of intensity equation (TIE) contrast methodology allows unlabled samples to be imaged and facilitates optimum fluorescent imaging through refined focusing. This system will support traditional IHC imaging as well as cyclic immunofluorescence studies. Contact the AMCF to learn more and/or schedule your reservation. Reference images from Zeiss.

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MILTENYI BIOTEC ULTRAMICROSCOPE II LIGHT SHEET FLUORESCENCE MICROSCOPE


Coming Soon: Installation Spring/Summer 2022!

Tissues are inherently three dimensional in nature, which makes imaging intact tissues a necessity for a more complete study in the relationship between structure and function and the system-level study of cellular mechanisms.

The Miltenyi Biotec (previously La Vision) Ultramicroscope II Light Sheet fluorescence microscope (LSFM) you can extend your existing fluorescent imaging into true 3D, large-scale volumetric imaging of intact tissues, organs, and small organisms. Unlike traditional laser scanning microscopy using parallel/identical excitation and collection paths, LSFM excites your sample in an orthogonal beam path. By setting the width, numerical aperture (NA), and Rayleigh length of illumination, users can optimize the shape of the light sheets for their specific sample size and imaging goals. A lower NA results in a broad field of view at the expense of a lower z-resolution and vice versa, with a full range of gradations in between. Using this approach LSFM can reduce out of focus light, capture larger fields of view and collect volumetric images 10’s to 100’s of times faster than traditional confocal microscopes.  Optimized volumetric imaging requires removal of light bending cellular components (mostly lipids) through a process called optical clearing. Clearing reagents differ. Light travels differently in different imaging media. The UltraMicroscope II’s onboard software allows users to choose between various preloaded clearing and imaging solutions to optimize imaging. Reference images from Miltenyi Biotech. Contact the AMCF to learn more and/or schedule your reservation.

To support your volumetric imaging studies, the AMCF also offers equipment for rapid, optimized tissue clearing using the X-Clarity System.

Tissue clearing is important for 3D imaging of tissues at single-cell resolutions. The X-CLARITY™ system simplifies and accelerates each step of the tissue clearing process using CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining /in situ-hybridization-compatible Tissue hYdrogel), a tissue clearing method developed by the Deisseroth lab at Stanford. Fixed tissues are embedded in a hydrogel matrix and lipids are actively extracted through electrophoresis to create a stable and optically transparent tissue-hydrogel hybrid that is chemically accessible for multiple rounds of antibody labeling and imaging. Native cytoarchitecture remains intact and endogenous fluorescence proteins are preserved for robust fluorescence imaging downstream. For LSFM imaging, samples should be optically cleared (X-clarity), labeled with antibodies (see kits below), then equilibrated in refractive index (RI) matched mounting solutions for imaging.

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HALO-EQUIPPED WORKSTATION

Coming Soon: Installation Spring/Summer 2022!

Maximize your imaging data sets using the premier image analysis platform, HALO (Indica Labs), to perform your quantitative tissue analyses. HALO reports morphological and multiplexed expression data on a cell-by-cell basis across entire tissue sections and maintains an interactive link between cell data and cell image. Click on any cell in the image and immediately see analysis outputs for that specific cell. Sorting and filtering capabilities allow the user to mine millions of cells while visually assessing corresponding cell populations. For example, sort cells according to biomarker intensity and immediately locate cells with highest intensity in the image. The AMCF has several HALO modules ready for use on the HALO image analysis workstation. Contact the AMCF to learn more and/or schedule your reservation.

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IMARIS-EQUIPPED WORKSTATION

Coming Soon: Software Updates Spring/Summer 2022!

IMARIS is the world’s leading Interactive Microscopy Image Analysis software focused on 3D and 4D imaging. The AMCF is equipped with the complete IMARIS for Core Facilities software package with focused image analysis, segmentation, and reconstruction options for cancer and neuroscience researchers, cell and developmental biologists. IMARIS can be used for any number of imaging studies but is anticipated to be particularly relevant for researchers using the Elyra PS1 super-resolution microscope and Ultramicroscope II light sheet fluorescence microscopes. Contact the AMCF to learn more and/or schedule your reservation. Top images, IMARIS, bottom super-resolution image, Casey et al. 2021.

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Acknowledgement

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