Instrumentation


LSR-II Flow Cytometer Click for configuration

LSR-II
  • Benchtop four-laser BD LSR-II with the ability to acquire up to 18 colors and analyze >20,000 events per second, providing an efficient acquisition of 1-20 million cells in a volume of 60 ul per minute.
  • This instrument provides non-sorting applications with high sensitivity and specificity due to the presence of lasers that improve signal-to-noise ratios.
  • The configuration of lasers and dichroic filters in the LSR-II allows for 13 different settings to capture fluorochromes and related reagents, including Q-dots and BD Horizon dyes, based on similar excitation and emission spectra.
  • In addition to the standard analysis of samples by forward/side scatters, this cytometer allows the detection of multiple fluorochromes at a time on the surface or within cells (including the detection of reagents that label DNA).
  • With the multiple lasers, detectors and fluorochrome options, the LSR-II will provide a wide variety of applications to analyze several markers in the same sample.

FACS Celesta with High Throughput Sampler (HTS) Click for configuration

FACS Celesta
  • 3-laser (blue, violet and read) benchtop flow cytometer with the capability to acquire 12 parameters.
  • Cytometer is coupled to a HTS device providing automated acquisition of samples from a multi-well plate.
  • Throughput modes allow for acquisition of 96-well plates in 15 – 44 minutes at flow rate of 1 µl/sec.

ImageStreamX-Imaging Flow Cytometer-ISX-MKII Click for configuration

Image Stream
  • 4-laser 12-channel Amnis ImageStream X imaging flow cytometer, provides the capabilities of the LSR-II but adds the ability to simultaneously visualize individual cells microscopically.
  • The specific localization of markers of interest within the cell can be quantified and monitored over time and compared among samples.
  • This equipment allows investigators to capture detailed multispectral images of cells at rates of up to 5,000 cells a minute using 10 fluorescent channels and two bright field channels.
  • By combining conventional flow cytometry and the functional insights of microscopy, this system allows the study of novel pathways such as internalization, co-localization and translocation in intracellular assays.
  • The system is equipped with a 96-well auto-sampler to allow for high-throughput sample processing and screening assays.
  • The myriad of applications with this instrument include microscopic observations as well as quantitative imaging that would be time consuming and laborious using standard microscopy.
  • By studying large numbers of cells or sub-cellular localization of markers, the imaging flow cytometer will provide an extensive range of novel applications.

FACS Aria Cell Sorter Click for configuration

FACS Aria

Equipped with four lasers, this instrument has the ability to detect 18 colors in a similar configuration to the LSR-II. To support research applications using human samples and samples that carry known pathogens or replication-defective retroviruses, the Core will provide access to sorting and cell analysis under a biosafety cabinet.


Zeiss 510 Meta Confocal

Zeiss 510

8 lasers lines:

  • Diode 405nm at 25mW
  • Tunable Argon 458/477/488/514nm
  • Helium-Neon 543nm
  • Helium-Neon 594nm
  • Helium-Neon 633nm
  • 2 Photomultiplier tubes (PMT)
  • 32-channel Meta spectral detector
    (polychromatic detector, 3rd PMT)

Imager Z1 upright WT fluorescence microscope
objectives:

  • 5X Plan-Apochromat/0.16 NA with DIC
  • 10X Plan-Neo/0.3 NA with DIC
  • 20X Plan-Apo/0.75 NA with DIC
  • 40X Plan-Neo/1.3 NA Oil with DIC
  • 63X Plan-Apo/1.4 NA Oil with DIC
  • 100X Plan-Apo 1.4 NA Oil with DIC

Zeiss 710 NLO 2P System – Live Cell Imaging

Zeiss 710

7 laser lines:

  • Diode 405nm
  • Diode 561 nm
  • Tunable Argon 458/477/488/514nm
  • Helium-Neon 561nm
  • Chameleon Ultra II Ti:Sa

Incubation Chamber

Inverted Observer Z1 WF fluorescence microscope
objectives:

  • 5X Plan-Apochromat/0.16 NA with DIC
  • 10X Plan-Neo/0.3 NA with DIC
  • 20X Plan-Apo/0.8 NA with DIC
  • 40X Plan-Neo/1.3 NA Oil with DIC
  • 40X C-Apochromat/1.2 Water
  • 63X Plan-Apo/1.4 NA Oil with DIC

Data Analysis

data analysis

Complementing available instrumentation, the Core houses three dedicated workstations for data analyses, available to investigators for training with flow cytometry software including FlowJo, IDEAS as well as applications for image analysis such as ImageJ and Imaris.

FlowJo is a user-friendly software with an integrated environment for viewing and analyzing flow cytometric data. The environment is presented as a workspace where samples are loaded and gates are easily generated along with statistical analyses and the generation of tabular and graphical layouts.

IDEAS software processes and analyzes the data gathered in the ImageStreamX system, allowing for the generation of histograms and scatter plots. Within this software, every dot on a scatter plot links to a cell’s imagery providing a virtual sorting capability. With this software the user can analyze images for any gated population using over 200 features providing statistical analysis for each one.

Imaris is an application that allows for visualization and analysis of microscopic data acquired with the Zen software. Among other applications, Imaris facilitates the visualization and production of high-quality images for presentation and storage by the availability of 8 different viewing options.

The core partners with the Research Computing Support Group to create a data acquisition, analysis and storage pipeline that ensures a streamlined and seamless workflow for researchers.


User Support Services

Our core provides the following services:

  • Access to state-of-the-art instrumentation for flow cytometry, confocal, and 2-photon imaging
  • Setting up and maintaining all instruments
  • Education and training in the use of core instrumentation
  • Assistance in the incorporation of flow cytometry, confocal microscopy, and live-cell imaging applications to researchers’ needs
  • Technical support for flow cytometry and confocal microscopy applications
  • Consultation on projects, data interpretation and analysis for manuscripts/grant proposals