Core Facilities

Confocal Microscope and Flow Cytometry Facility

Wade J. Sigurdson, PhD, has expertise on instrumentation including the Leica TCS SP8 confocal microscope system.

Our instrument laboratory provides training and consultation services for researchers whose work involves flow cytometry, confocal microscopy and gel-documentation. We work closely with other core facilities to provide comprehensive and integrative services to solve the imaging and cytometry challenges presented by cutting-edge research programs.   

Flow Cytometry (FACS) Instrumentation

Our facility hosts two analytical flow cytometers:

  • BD FACS Fortessa Special Order Product (SORP) analytical flow cytometer
    • Five excitation laser lines, 355nm, 405nm, 488nm, 561, 640nm
    • 20 parameter detection with 18 colors
    • Ready to accept High Throughput Sampler option
  • BD FACS Celesta analytical flow cytometer
    • Three excitation laser lines, 405nm, 488nm, 640nm
    • 14 parameter detection with 12 colors
    • High Throughput Sampler option installed to accept multi-well plates
    • Can be configured to perform multiplex analyses using the cytometric bead array technique

Both of these instruments are operated by BD’s DiVA software platform for acquisition control. We also offer a networked version of DeNovo’s FCSExpress cytometry analysis program.

Our BD FACS Aria Fusion cell sorting flow cytometer has these features:

  • Four excitation laser lines, 405nm, 488nm, 561, 640nm
  • 18 parameter detection, 16 colors
  • Cell sorting at 20,000 cells/sec into either 4 collection tubes or into multi-well plates
  • Sorter is contained within a Biological Safety cabinet providing BSL-2 enhanced biosafety protection
  • Cell sorting is done aseptically
  • A laminar air flow hood and incubator are provided for pre- and post- sort cell maintenance procedures

We have a Zeiss K-Elispot automated imaging analysis system.

Ray Kelleher.

Raymond Kelleher, PhD, can assist you with flow cytometers such as the BD FACS Fortessa.

Microscopy Instrumentation

Confocal Microscopes

Both of our confocal microscopes are capable of super-resolution imaging, where diffraction limited resolution is exceeded by a variety of techniques.

Leica TCS SP8 confocal microscope system, installed on a Leica DMi 8 inverted stand:

  • Excitation lasers
    • White Light Laser (470nm-670nm) with 200 available lines (all pulsed)
    • 460nm pulsed laser (suitable for CFP excitation)
    • 405nm diode laser (suitable for UV excited dyes such as DAPI)
    • Excitation lines selected by an acoustic optical beam splitter (no dichoric mirrors
    • 8kHz Resonant scanning mode can produce 28 frames/second 512x512 images
  • Spectral Emission detectors (no emission filters)
    • Two PMT
    • Two standard hybrid detectors (high sensitivity photocathode and APD)
    • One low noise single molecule hybrid detector
    • Emission bandpass adjustable to match fluorochrome emission spectra
  • 3D Stimulated Emission Depletion (STED) super-resolution
    • Lateral resolution less than 50nm
    • X, Y and Y resolution at least 120nm
    • Three depletion lasers 592nm, 660nm and 775nm permitting super-resolution imaging of samples with multiple fluorochromes
  • Leica’s new FALCON fluorescence life-time imaging (FLIM) system
    • Using the pulsed white light and 440nm lasers
    • And hybrid detectors, fluorescence life times can be easily and very quickly determined
    • FLIM can be used as a contrasting method for samples where spectrally identical dyes have been used, eg. GFP and Alexa 488
  • Digital Light Sheet (DLS) imaging mode
    • Using the resonant scanner system and the confocal excitation lasers, specific illumination objectives and mirror-caps attached to the imaging objective light sheet imaging of large (~5mm working distance) and smaller (2.5mm working distance) samples
    • 3 illumination objectives and 3 imaging objectives (5x, 10x, 25x water immersion)
    • High-speed sCMOS camera
  • Leica Hyvolution acquisition and deconvolution package
    • Employing over-sampling and Huygens deconvolution confocal datasets can be processed to improve resolution in x, y and z and increase image signal to noise
    • Later resolution is increased to ~140nm and z axis resolution 2 fold
    • Deconvolution processing is hardware accelerated to reduce processing time
    • Huygens deconvolution software and Leica LAS X are integrated making Hyvolution processing seamless

VisiTech VT-I iSIM swept-field super-resolution confocal microscope system:

  • Hosted by a Leica DMi 6000B inverted fluorescence microscope
  • Laser engine includes 405nm, 488nm, 568nm, 647nm lines for excitation of up to 4 fluorochromes simultaneously
  • 100x/1.47 TIRF oil immersion objective
  • Camera link sCMOS camera allows image acquisition at up to 400 frames/sec
  • Two-fold super-resolution improvement after Hyvolution GPU accelerated deconvolution

Wide-field Fluorescence Microscopes

We have three wide-field fluorescence microscopes, all of which are motorized and computer controlled to ease user operation and allow complex image acquisition protocols.

Leica DMi 8 inverted fluorescence microscope for live cell imaging and fast acquisition:

  • Equipped with objectives (10x, 20x, 40x, 63x oil immersion) suitable for imaging multi-well plates, culture dishes and microscope slides in bright-field, phase contrast, differential interference contrast (DIC) and fluorescence in high resolution (40x/1.1 water immersion with motorized correction collar)
  • 16 LED fluorescence illuminator for very fast excitation switching combined with a 4 color bandpass filter cube
  • sCMOS Leica camera with high-speed camera link connection to the acquisition computer
  • Environmental control of sample via a PeCon heated microscope housing and a Tokaihit stage top incubator with control of temperature and CO2
  • Leica LAS X software package includes full control of microscope acquisition parameters including montaging and image stitching, z stacks and templates for various sample holders
  • Autoquant blind deconvolution software integrated into LAS X for seamless deconvolution image processing of 3D datasets

Leica DMi 8 inverted microscope for bright-field, phase contrast, DIC and fluorescence imaging:

  • Similar to the inverted fluorenscence microscope except for the camera-link high speed imaging option and LED excitation source
  • Equipped with 5x, 10x, 20x, 40x and 63x oil immersion objectives
  • Leica LAS X software package includes full control of microscope acquisition parameters including montaging and image stitching, z stacks and templates for various sample holders

Leica DM 6B upright microscope for bright field, DIC and fluorescence imaging:

  • Objectives include 5x, 10x, 20x, 40x oil and 63x oil immersion
  • Includes the Lumencore excitation light source, Leica sCMOS camera for low-light imaging and a Leica high-resolution color camera for chromogenic samples
  • Autoquant blind deconvolution is included and integrated into LAS X

Leica LAS X software package includes full control of microscope acquisition parameters.

Ancillary Instrumentation

BioTek Cytation 1 multi-function plate reader and microscope:

  • Plate reader capable of reading absorbance, fluorescence and luminescence
  • Bright-field and fluorescence imaging of multi-well plates and microscope slides
    • 4x and 20x objectives
    • DAPI, Green, Orange, Texas Red and far red fluorescence
  • Incubation capable with temperature and CO2 control for long term imaging and reading experiments

Bio-Rad Chemi-Doc MP multimode imager:

  • Imager for Western Blots and electrophoretic gels
  • Colorimetric, Luminescence and Fluorescence imaging including Infrared dyes

Bio-Rad Gel Doc EZ gel imager:

  • Commassie, Ethidium bromide, Sybr dye stained gels

Bio-Rad Real Time qPCR thermocyclers:

  • CFX-95 Touch Multi-plex (5 color channels) qPCR
  • CFX-95 Connect Multi-plex (5 color channels) qPCR

Two Thermo-Fisher NanoDrop One spectrophotometers.

GE Amersham Typhoon IP/IR scanner for radio-labelled and infrared fluorescence samples.

Leica AS/LMD laser microdissection/microcapture system.

Location

955 Main Street
Rooms 3130 and 3135
Buffalo, NY 14203

Contact Information

Flow Cytometry Administrator

Kelleher, Raymond

Raymond Kelleher, PhD

Research Associate Professor

Jacobs School of Medicine and Biomedical Sciences, 955 MainStreet, Room 5122 Buffalo, NY 14203

Phone: (716) 829-2558

Email: rjk6@buffalo.edu

Confocal Microscopy Facility Director

Wade J. Sigurdson, PhD.

Wade J. Sigurdson, PhD

Director

Confocal Microscope and Flow Cytometry Facility

955 Main Street, Suite 3130/3135, Office 3134, Buffalo, NY 14203-1121

Phone: (716) 829-3589; Fax: (716) 829-3589

Email: wjs@buffalo.edu

Make online reservations through our scheduling system. Contact the director for login information.