About Us

The Flow Cytometry & Confocal Microscopy Facility provides investigators with access to sophisticated instruments designed to detect and measure fluorescent light emission. The flow cytometers distinguish cells or particles based on size, internal complexity, and fluorescent signals. Cells or particles can also be sorted to obtain pure populations for further analysis or subsequent culture. Available imaging techniques include widefield, laser scanning confocal, spinning disk confocal, TIRF, and multiphoton confocal microscopy.

From left to right: Co-Facility Heads Joseph LoTurco, Michael Lynes, and Adam Zweifach; Facility Scientist Carol Norris; Co-Facility Heads David Knecht and Lawrence Silbart

Services and Instrumentation

Please contact Facility Scientist Carol Norris for additional information or consultation: carol.norris@uconn.edu, (860) 486-3080.

Flow Cytometry

BD Biosciences FACSAria II Fluorescence-activated Cell Sorter

• Three lasers provide excitation at 407, 488, and 633 nm for analysis of up to 10 fluorescence channels plus forward and side scatter
• Digital electronics
• Sort up to four populations simultaneously

BD FACSCaliber Flow Cytometer

• Two lasers provide excitation at 488 and 633nm for analysis in four fluorescence channels plus forward and side scatter
• Straightforward operation allows independent use by researchers
• Offline computer with FlowJo analysis software

FlowJo Analysis Workstation

• Post-acquisition compensation to correct cross-talk between fluorescence channels
• Multiple graphing options
• Algorithms for analysis of cell cycle and cell proliferation data

Facility Scientist Dr. Carol Norris at the FACSCalibur

Imaging

Nikon A1R Spectral Confocal Microscope

• Excitation wavelengths: 405, 457, 476, 488, 514, 561, 640 nm
• High efficiency detectors
• 32 channel spectral detector for separation of overlapping signals
• FRET, FRAP, photoactivation, photoconversion
• Computer-controlled x-y stage
• Incubation chamber for control of temperature and CO2.
• Nikon Perfect Focus system

Facility Co-head Dr. David Knecht and Nikon Instruments representative Mr. Brad Coyle demonstrate the use of the Nikon A1R for live cell confocal imaging.

Andor Confocal & TIRF Microscope

• High speed 4D confocal imaging or TIRF microscopy with EMCCD camera
• Incubation chamber maintains live cells at 37?C and 5-10% CO2
• Four lasers for excitation at 405, 488, 561, and 633 nm
• Nikon Perfect Focus system

Zeiss LSM 5 Multiphoton Confocal

(Contact Facility Head Joseph LoTurco)

• System configured for intravital imaging of fluorescently labeled cells
• Zeiss Axoskop 2 FS upright scope with Sutter mechanical stage to accommodate most small animal preparations
• Chameleon Ultra II pulsed laser for multiphoton excitation of eGFP or mRFP

Leica TCS SP2 Laser Scanning Confocal

• Excitation wavelengths: 458, 476, 488, 543, 633 nm
• Three fluorescence detectors plus transmitted light
• Reflection mode, xzy imaging, wavelength scanning, time lapse, FRET

Zeiss Axiovert Widefield Microscope

• Filter sets for: DAPI, FITC, TRITC, GFP, Cy5, CFP-YFP FRET, Texas Red
• Computer-controlled operation for complex time lapse imaging
• High sensitivity Hamamatsu CCD camera and Qimaging color camera

Available Reagents

FCCM has a small supply of reagents available at cost for pilot experiments. Contact the Facility Scientist for details.

Training and Education

FCCM Staff provide training and consultation in the use of facility instruments. Consultations are free but prospective users must schedule an appointment.

Cost for Services

Current pricing can obtained by contacting the Facility Scientist.

All current rates are subject to periodic review and change.