Equipment & Pricing

Training

Hands-On Training is required of all new users after completion of the Introduction to Flow Cytometry class and and is instrument specific.  Training covers instrument operation, data collection, biosafety, troubleshooting, and general maintenance. Reserve 1.5 hours on the instrument of choice for Hands-On Training on the Flow Cytometers. Fluorescent Beads will be provided for training. Please do not bring cell samples.

For training on the ImageStream, please reserve 2.5 to 3 hours.  Training works best on this instrument with actual samples, so please bring experimental samples for your first training on the ImageStream.  This will provide an opportunity to assess sample prep, suggest steps for optimizing your assay going forward, and a data set to use for learning the IDEAS image cytometry data analysis software.

Training appointments are available at the following times:

  • ThermoFisher Attune* (Lightning):  Tuesdays, 11am-3pm
  • ThermoFisher Attune* (Thunder):  Thursdays, 11am-3pm
  • BD LSRII:  Wednesdays, 11am-3pm
  • BD LSR Fortessa:  Reserve BD LSRII, Wednesdays, 11am-3pm
  • ImageStream:  Fridays, 11am-3pm
  • Aurora: Mondays, 11am-3pm

After completion of Hands-On Training, users will schedule Assisted appointment(s) with actual experimental samples.  These Assisted appointments will focus on optimizing instrument settings for their specific experiments. Training will be complete when both the user and Flow Lab staff feel the user is competent to operate the instrument(s) independently.  Users will be checked off as trained in iLab for individual instrument(s) allowing user access to reserve instrument(s) outside of the reserved training times.

*Those eligible for use of the Attune in the SAIRF or Peds should reserve training slots on Thunder or Lightning. Those wanting to use the Attune V6 at Biotech should do their Hands-On Training on Thunder or Lightning and then work with the Flow Lab staff to schedule training time with actual samples on the Attune V6 instrument to complete the training program and optimize settings for their individual experiments.

Equipment

Cytek Aurora Spectral Flow Cytometer

Image of Cytek Aurora Spectral Cytometer

The Cytek Aurora is a 5 Laser Spectral Cytometer, capable analyzing 40+ color cytometry samples.  This instrument captures the bulk of the emission spectra of each fluorochrome, allowing for fluorochrome combinations not possible on traditional flow cytometers, thus increasing the number of fluorochromes possible per panel.  This instrument is equipped with 5 lasers (355nm UV, 405nm Violet, 488nm Blue, 561nm Yellow-Green, and 640nm Red). It has three light scatter channels, Forward Scatter and Side Scatter off both the Blue and Violet Lasers, and 64 fluorescence channels. Samples can be acquired from traditional 12x75mm flow tubes as well as from 96 well plates.

Contact the Flow Lab to set up a personalized consultation or training session at uwflow@uwcarbone.wisc.edu

Aurora 5 Laser Fluorochrome Guide

Cytek Full Spectrum Viewer

OMIP‐069: Forty‐Color Full Spectrum Flow Cytometry Panel for Deep Immunophenotyping of Major Cell Subsets in Human Peripheral Blood

  • Two ThermoFisher Attune Next Flow Cytometers: “Thunder” and “Lightning”
    UW Flow Cytometry Attune NxTLocation: 7016 WIMR

    The Attune NxT is a benchtop analysis cytometer with 4 lasers (488, 405, 561, and 633nm). Capable of collecting up to 16 parameters; 2 scatter and 14 fluorescent, the optical benches are user configurable to accommodate unusual fluorochromes and unique combinations. These instruments use acoustic focusing to center cells in the stream, allowing for significantly increased acquisition speeds (up to 1000uL per minute) with little to no loss in resolution. They are also equipped with plate auto-samplers for regular and deep-well 96 and 384 well plates.

    Attune Optical Bench Map (pdf opens in new tab)

  • ThermoFisher Attune NxT V6 Flow Cytometer: “River”

    ThermoFischer Attune Nxt V6 "River,"

    Location 2360 Biotech – Available Spring 2019

    The Attune NxT is a benchtop analysis cytometer with 4 lasers (488, 405, 561, and 633nm). Capable of collecting up to 16 parameters; 2 scatter and 14 fluorescent, the optical benches are user configurable to accommodate unusual fluorochromes and unique combinations. These instruments use acoustic focusing to center cells in the stream, allowing for significantly increased acquisition speeds (up to 1000uL per minute) with little to no loss in resolution. They are also equipped with plate auto-samplers for regular and deep-well 96 and 384 well plates. The V6 model is configured with 6 channels off the violet laser, allowing expanded usage of the available violet-excitable dyes including the SuperBright and Brilliant Violet dye series.

    A versatile instrument, the Attune V6 can be used for multi-color immunophenotyping, automated high-throughput screening of samples in plates (i.e., assessment of transfection efficiency), as well as common flow cytometry assays such as cell cycle, apoptosis & cell death, phosphoproteins, mRNA, intracellular cytokines and transfection factors, proliferation, fluorescent proteins, et. al.

    Attune V6 Optical Bench Map (pdf opens in new tab)

  • ThermoFisher Attune Next Flow Cytometer for Analysis of Radiotherapy Samples
    UW Flow Cytometry Attune NxTLocation: WIMR B1334

    Managed by the UWCCC Flow Lab and located in the UWCCC SAIRF, this instrument is reserved for the analysis of radiotherapy samples. Users must have completed the Flow Lab’s training program, and Radiation Safety Training in order to use this instrument.

    The Attune NxT is a benchtop analysis cytometer with 4 lasers (488, 405, 561, and 633nm). Capable of collecting up to 16 parameters; 2 scatter and 14 fluorescent, the optical benches are user configurable to accommodate unusual fluorochromes and unique combinations. These instruments use acoustic focusing to center cells in the stream, allowing for significantly increased acquisition speeds (up to 1000uL per minute) with little to no loss in resolution. They are also equipped with plate auto-samplers for regular and deep-well 96 and 384 well plates.

    Attune Optical Bench Map (pdf opens in new tab)

  • ThermoFisher Attune NxT “Peds”

    ThermoFischer Attune Nxt V6 "River,"

    Location WIMR 4048

    Acquired through generous gifts from the Lion’s Club and the MACC Fund, this instrument is reserved for Pediatric Cancer Research Labs.  Please Contact the Flow Lab for more information.

    The Attune NxT is a benchtop analysis cytometer with 4 lasers (488, 405, 561, and 633nm). Capable of collecting up to 16 parameters; 2 scatter and 14 fluorescent, the optical benches are user configurable to accommodate unusual fluorochromes and unique combinations. These instruments use acoustic focusing to center cells in the stream, allowing for significantly increased acquisition speeds (up to 1000uL per minute) with little to no loss in resolution. They are also equipped with plate auto-samplers for regular and deep-well 96 and 384 well plates.

    Attune Optical Bench Map (pdf opens in new tab)

BD LSRII

The BD LSRII is reaching end of life and will be retired on December 31, 2022.  Please contact the Flow Lab at uwflow@uwcarbone.wisc.edu if you have an ongoing experiment that extends beyond the end of 2022 to work with the staff to create a plan for seamless transition to another instrument.

UW Flow Cytometry BD LSRII

Location: 7016 WIMR

The LSRII is a 5 laser (355nm, 405nm, 488nm, 561nm, and 640nm) 20 parameter (2 scatter and 18 fluorescence) benchtop analysis cytometer.  The optical bench is user configurable for detection of less common fluorochromes and unique combinations.

LSR Optical Bench Map (pdf opens in new tab)

BD LSR Fortessa

UW Flow BD LSRFortessaLocation: 7016 WIMR

The LSRII is a 5 laser (355nm, 405nm, 488nm, 561nm, and 640nm) 20 parameter (2 scatter and 18 fluorescence) benchtop analysis cytometer.  The optical bench is user configurable for detection of less common fluorochromes and unique combinations.  The Fortessa also has a High-Throughput Sampler for automated sampling of 96 and 384 well plates.

Fortessa Optical Bench Map (pdf opens in new tab)

ImageStream

UW Flow Cytometry ImageStreamX MarkIILocation: 7016 WIMR

The ImageStream MarkII is an imaging cytometer for cells in suspension. It has high resolution CCD cameras instead of photomultiplier tubes for detection and is equipped with 20x, 40x and 60x objectives, six detection channels and two lasers (488nm and 642nm).

ImageStream Optical Bench Map (pdf opens in new tab)

  • BD FACSAria with Biosafety Cabinet “Jill” in 7017 WIMR

    UW Flow Cytometry BD FACSAriaII "Jill"

    Cell sorting flow cytometer capable of sorting up to 4 populations simultaneously into tubes, single and multiple cell deposition into 6 – 384 well plates and other custom devices.  Equipped with 5 lasers:  488nm, 405nm, 640nm, 532nm, and 355nm, 16 fluorescence detection channels, and ability to customize emission filters for specific fluorochromes. Contained in custom biological safety cabinet for sorting up to BSL-2 single cell suspensions.
    Aria “Jill” Optical Bench Map (pdf opens in new tab)

  • BD FACSAria with Biosafety Cabinet “Jack” in 7016 WIMR
    Cell sorting flow cytometer BD FACSAria with Biosafety Cabinet "Jack"
    BD FACSAria with Biosafety Cabinet “Jack”

    Cell sorting flow cytometer capable of sorting up to 4 populations simultaneously into tubes, single and multiple cell deposition into 6 – 384 well plates and other custom devices. Equipped with 4 lasers: 488nm, 405nm, 640nm, and 561nm, 13 fluorescence detection channels, and ability to customize emission filters for specific fluorochromes.  Contained in custom biological safety cabinet for sorting up to BSL-2 single cell suspensions.

    Aria “Jack” Optical Bench Map (pdf opens in new tab)

  • BD FACSAria in BioBubble “Jayne” in 2360 Biotech
    BD FACSAria "Jayne". Cell sorter equipped with 5 lasers and 18 fluorescence detection channels.
    BD FACSAria “Jayne”

    Installed in March of 2019, this cell sorter is equipped with 5 lasers (488nm, 405nm, 633nm, 561nm, and 355nm) and 18 fluorescence detection channels. This instrument is contained in a BioBubble negative pressure containment unit and is strictly limited to BSL-2 or lower samples only. It is capable of sorting up to 4 populations simultaneously into tubes, single and multiple cell deposition into 6 – 384 well plates and other custom devices.

    This Aria has been placed in the UW Biotechnology Center to support Single Cell Genomics pipelines as well as to provide cell sorting to central campus research labs.  Located in the same building as the following cores; Gene Expression Center, DNA Sequencing, Genome Editing & Animal Models, and the Bioinformatics Resource Center, this instrument is uniquely positioned to provide highly purified cell subpopulations and single cell per well deposition into up to 384 well plates for downstream genomics assays.

    Aria “Jayne” Optical Bench Map (pdf opens in new tab)

Scheduling – The Flow Lab may adjust requested start times for sort appointments to optimize available instrument time.

Biosafety and Cell Sorting

Anything sorted in the Flow Lab MUST be described on your Lab’s Biological safety protocol.  Cell sorting creates aerosols and additional biological safety precautions are required.  The Flow Lab is able to sort samples of BSL-2 and below.

Flow Lab Hood Certification Dates

  • Baker BioProtect Aria “Jill” Cell Sorter (7018B WIMR), SN 120708, Cert. Date: 12/03/18
  • Baker BioProtect Aria “Jack” Cell Sorter (7018B WIMR), SN 108121, Cert. Date: 12/03/18
  • BioBubble Aria “Jayne” Cell Sorter (2360 Biotech), SN PU2525-0159, Cert. Date: 07/15/20
  • 4′ Baker SteriGuardIII Advance (7018E WIMR), SN 93707, Cert. Date: 12/03/18
  • 6′ Baler SteroGuardIII Advance (7018E WIMR), SN 95206, Cert. Date: 12/03/18
  • 4′ Baker SteriGuardIII Advance (2360 Biotech), SN 90115, Cert. Date: 07/13/20

Sony MA900 Cell Sorter

Single Cell Deposition by Sony MA900
Single Cell Deposition into 384-well plate using beads and Horseradish Peroxidase indicator on Sony MA900 Cell Sorter

The Sony MA900 Multi-Application Cell Sorter with sample and collection devices.

 

The Sony MA900 is a highly automated cell sorter allowing for end-user self-operation and we will be training researchers to operate this instrument independently.  This will allow for more flexible scheduling, and will be available to trained researchers after-hours and on weekends to accommodate unusual time points, long pre-processing workflows, and clinical research samples with later arrival times.

 

Training is composed of 2 sessions:

  • AM Session:  9:30-11:30am – Start-up & QC Training
  • PM Session: 1:30-3:30pm – Cell Sorting & Shutdown Training.  Customers should bring stained cells for sorting. Training will include setting up the sort, adjusting sensitivity, compensation, sorting, saving a template, rigor and reproducibility considerations, and instrument shutdown.
  • Staff is available for additional training/assistance as needed.  Please email the Flow Lab at uwflow@uwcarbone.wisc.edu if you need staff assistance during a self-run reservation during set-up, or for a new assay.

The MA900 is capable of sorting up to 4 populations simultaneously into tubes, single and multiple cell deposition into 6 – 384 well plates and other custom devices, including PCR plates.  Equipped with 4 lasers arranged in co-linear pairs:  488nm & 561nm, and, 405nm and 634nm, 12 fluorescence detection channels, and ability to customize emission filters for specific fluorochromes. Contained in custom biological safety cabinet for sorting up to BSL-2 single cell suspensions.

Sony MA900 Optical Bench

Sony MA900 Fluorochrome and Filter Guide

Sony MA900 Manufacturer Webpage

Located at our WIMR location, the Nexcelom Cellometer Auto T4 Cell Viability Counter is a PC-based instrument for one-step 4x bright field image capture and analysis. This state-of-the-art cell counter performs count, size, and Trypan Blue viability analyses that are 10x faster and more accurate than manual counting. This instrument is ideal for cell lines and purified primary cells, including MCF-7 breast cancer cells, PC3 prostate cancer cells, and 1600 other cell types, due to its advanced software that assesses clumpy cells and omits debris. This is a slide based cell counter and requires Nexcelom counting slides. There are 2 tests per slide and the slides can be bought individually or in packs of 75 from the Flow Lab.

Offline data analysis computers are available to laboratory users only and are located at both the 7th floor WIMR location and at the Biotech Center (opening Spring 2019). Computers are loaded with a variety of analysis packages including FlowJo,  FCS Express, ModFit, WinList, and IDEAS ImageStream analysis software, Microsoft Office, and Adobe Illustrator.
There is no charge for the use of these computers and are for the exclusive use for the analysis of data generated at this laboratory. Computers are available on a first-come, first-served walkup basis for users of the facility.

Pricing

Hourly Rates – Effective January 1, 2022

Service
UWCCC Member
Non-Member
External Academic
Off Campus
Attune Training/Assisted $100.00 $142.86 $222.14 $295.45
Attune Self $60.00 $85.71 $133.29 $177.27
Aurora Training/Assisted $100.00 $142.86 $222.14 $295.45
Aurora Self $60.00 $85.71 $133.29 $177.27
LSR & Fortessa Training/Assisted $100.00 $142.86 $222.14 $295.45
LSR & Fortessa Self $60.00 $85.71 $133.29 $177.27
ImageStream Training/Assisted $100.00 $142.86 $222.14 $295.45
ImageStream Self $60.00 $85.71 $133.29 $177.27
Staff Assistance (Analysis Cytometers)* $40.00 $57.14 $88.86 $118.18
LSR & Fortessa Weekend Use Training $450.00 $642.86 NA NA
Staff Assisted De-Clog Fee* $110.00 $157.14 $244.36 $325.00
Cell Sorting (Aria & Sony MA900) Training/Assisted $105.00 $150.00 $233.25 $372.27
Cell Sorting (Aria & Sony MA900) Self $81.00 $115.71 $179.94 $310.22
Sorter Set-up Fee $52.50 $75.00 $116.63 $186.13
Aria Weekend Use Training 472.50 $675.00 NA NA
Consultation (Discussion Only) No Charge $30.00 $46.65 $62.04
Sort Consultation (Req’d before 1st Sort) No Charge No Charge $46.65 $62.04
Assay Services $110.00 $157.14 $244.36 $325.00
Aria Training and Weekend Usage requires sustained sorting needs of at least once per week.
Assay Services include: Protocol Optimization & Development, Panel Design, Data Analysis, Data Reporting & Figure Presentation.
*15 minute minimum charge

Per Usage Fees

Service
UWCCC Member
Non-Member
External Academic
Off Campus
Lecture Series (Req’d before using instrumentation) $75.00 $107.04 $169.94 $255.00
Cellometer Cell Counter Customer purchases counting slides