CCAM Microscopy User Facility

CCAM Microscopy User Facility

The Center for Cell Analysis and Modeling (CCAM) Microscopy User Facility provides the UConn research community and other academic and industrial institutes, access to its state-of-the-art equipment for quantitative fluorescence imaging applications. The facility offers access to high end confocal microscopes, one integrated with a two-photon laser, and a light sheet microscope for imaging large, optically cleared samples and for the long-term observation of live biological specimens. CCAM offers its users access to quantitative software, and provides technical support, consultation, and opportunities for collaborative projects.

Contacts

pacman

Yi Wu, Ph.D.

Director, CCAM Microscopy User Facility

Associate Professor of Genetics and Genome Sciences

yiwu@uchc.edu
860.679.1773
pacman

Susan Staurovsky

Lab Manager
staurovsky@uchc.edu
860.679.4686

Instrumentation

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Mouse retina pigments epithelium cells/choroid wholemount with tight junction (anti- ZO1, Green) and ROS stress levels (anti-8OHdG, purple). (Y. Jiang/Fong Lab)
Mouse retina pigments epithelium cells/choroid wholemount with tight junction (anti- ZO1, Green) and ROS stress levels (anti-8OHdG, purple). (Y. Jiang/Fong Lab)

ELYRA 7 SIM2

Structured illumination microscopy, SIM, is a widefield technique that greatly increases the diffraction-limited resolution of the light microscope, ~ 200 nm, to super-resolution, ~ 100 nm. SIM uses a periodic grating that is projected onto the image plane of the microscope. Software is then used to analyze the resulting Moiré pattern, and through deconvolution, a super-resolution image is obtained.  The Zeiss Elyra 7 SIMuses an advanced reconstruction algorithm that improves the SIM resolution to less than 100 nm, along with quick image acquisition and compatibility for live-cell applications.

The Elyra is capable of other high-resolution techniques such as TIRF, PALM, and STORM. Samples live and fixed, up to ~ 150 µm thickness can be viewed on the Elyra 7 SIM.

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Zeiss LSM 880 Confocal Microscope

The 880 is mounted on an Axio Observer Z1 with automated xyz control. The system is equipped with a spectral 32-channel QUASAR GaAsp detector allowing for spectral imaging and maximum sensitivity photon counting. This detector is effective from 410 - 696 nm. There are two adjacent PMT's, effective range 415 - 735 nm, and a transmitted light detector. The accompanying incubation system, which encloses the microscope, offers stable environmental conditions with temperature, humidity and carbon dioxide control.

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3T3 cells stained with MitoTracker Red and Alexa Fluor 488 phallodin. (CCAM)
3T3 cells stained with MitoTracker Red and Alexa Fluor 488 phallodin. (CCAM)

Zeiss Observer Z1 - TIRF

The Zeiss Observer Z1 is a motorized, inverted microscope fitted with a stage-top incubation system for live cell imaging, providing heating, humidification, and carbon dioxide. The system is equipped with a 5.5-megapixel, Andor Neo sCMOS camera. The scope is capable of phase, DIC, and fluorescence microscopy. The Lumencor Spectra 7, 6 LED light source is currently configured for imaging DAPI, GFP, Rhodamine, and Cy5. Objective choices range from 10x through 63x.

An advanced imaging feature of this system is its capability to perform TIRF using 488 nm, 514 nm, and 561 nm wavelengths for fluorescent indicators such as GFP,  YFP, and RFP using the 100x/1.45 alpha-Plan Fluar oil immersion objective.

The microscope is controlled via MicroManager, a plugin from FIJI, enabling it to acquire single-channel or multi-channel images as well as time series with multiple position acquisition.

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IMAGING01

This CCAM user computer is a Windows 7 computer with MetaMorph, Photoshop, Office, Zeiss LSM and Zen Image Browsers. All the software is available, with prior approval, via remote access.

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Zeiss LSM 780 Confocal Microscope

The 780 is a combined confocal/FCS/NLO system, mounted on an inverted Axio Observer Z1. The 780 employs the spectral 32-channel QUASAR GaAsp detector allowing for spectral imaging, maximum sensitivity photon counting, and Fluorescence Correlation Spectroscopy (FCS) as well as two adjacent PMTs.  The GaAsp detector has a detection range of 410 - 696 nm while the PMTs have a range from 415 - 735 nm. The accompanying incubation system, which encloses the microscope, offers stable environmental conditions with temperature, humidity and carbon dioxide control. This microscope is also equipped with a Becker-Hinkl Fluorescence Lifetime Imaging Microscopy (FLIM) detector on a nondescanned detection port, particularly useful for detection of fluorescence resonance energy transfer (FRET) based probes.

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Zeiss LightSheet

The Zeiss LightSheet is an advanced fluorescent microscope that allows you to image optical sections of large, live or fixed samples with virtually no bleaching or phototoxicity to the specimen.  The optics are equipped for water-based or cleared specimens and dual sCMOS cooled cameras to provide high sensitivity, rapid, 3D imaging of live or fixed thick specimens. The extremely fast acquisition speeds allow you to capture large amounts of data within a fraction of the time compared to other imaging techniques.

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3 Day experiment, captured on this setup, observing 3T3 cells, in four different positions, was visualized using transmitted light and maintained with 5% carbon dioxide and 37-degree environmental conditions.

AxioVert 200M

The Zeiss AxioVert 200M is a motorized, inverted microscope fitted with an incubation system for live cell imaging, providing heating, humidification, and carbon dioxide environmental conditions. The system is equipped with a Hamamatsu ORCA  Digital CCD camera, with 70% quantum efficiency and 1344 (H) x 1024 (V) pixels. The scope is capable of phase and DIC microscopy and is fitted with a Lumencor Spectra 7, 6 LED light source for imaging with CFP, GFP, OFP/mCherry, and Cy5. MetaMorph software is used for the acquisition and analysis of data, including the ability to capture long-term, time series with multiple positions.

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Imaris

Imaris is a 64 bit computer with ImarisSurpass 3-D rendering and analysis software, MetaMorph, Photoshop, and Zeiss Zen Image Browser.

 

All software is available, with prior approval,  via remote access.

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Fluorolog

The Horiba-Jobin Yvon Fluorolog is a dual-emission spectrofluorometer. The visible detector has an emission range of 290 nm - 850 nm while the IR detector has an emission range of 850 nm - 1600 nm. The system is capable of emission scans, where the excitation wavelength is fixed, while the emission monochromator scans the sample, and excitation scans, where the emission wavelength is fixed, while the excitation monochromator changes the incident light. The sample chamber can also be temperature-controlled.

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Software

CCAM offers a wide variety of software ranging from applications for analyzing FRAP and other image data to tools for generating models and model parameters, including

    • Virtual FRAP Tool
    • SpringSaLaD
    • BioNetGen
    • Microfilament Detector
    • SyBil/SBPax

Services and Rates

Campus Address

Academic Research Building (ARB)
E6011

Cell & Genome Sciences Building (CGSB)
Imaging Suite R1510
UConn Health

Mailing Address

400 Farmington Avenue
Farmington, CT 06030-6406

263 Farmington Avenue
Farmington, CT 06030