Getting Started
Please review Using the Facility before making an appointment.
New users must attend the Intro to Flow Cytometry lecture series before signing up for any training on any cytometer. Training on the microscope does not require the lecture series.
After attending the lecture series, make an appointment in iLab for 1.5 hours on the instrument(s) to be used. New users will have access to only the specified Assisted/Training times listed below until training is completed. Please contact the Flow Lab at uwflow@uwcarbone.wisc.edu if you need to schedule Assisted/Training outside specified times.
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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, 12-3pm
- ThermoFisher Attune (Thunder): Thursdays, 12-3pm
- BD LSRII: Wednesdays, 12-3pm
- BD LSR Fortessa: Reserve BD LSRII, Wednesdays, 12-3pm
- MACSQuant10: Fridays, 12-3pm
- ImageStream: Mondays, 12-3pm
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.
Equipment
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Analysis Computers
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.
Benchtop Analyzers
The benchtop instruments are the Attunes, LSRII, Fortessa, MACSQuant 10, and ImageStream.
New users must first attend the Introduction to Flow Cytometry lecture series.
Use iLab to make appointments.
When signing up for appointments, ensure all fluorochromes to be used can be excited with the lasers listed.
Before using an instrument for the first time, schedule a 1.5 hour training appointment during the Assisted/Training times listed above. Please contact the Flow Lab at uwflow@uwcarbone.wisc.edu if you need to schedule Assisted or Training outside the listed times.
The instruments are available to trained users for self-run appointments. Staff-assistance is available to infrequent users and those who need or want additional assay-specific instruction.
All users operating cytometers after staff hours (6pm to 8am) or at the satellite facility are responsible for shutting down the cytometer after use. Please contact staff with any questions.
Instrument Maps (links open pdf in new tab):
- Attune Map – Thunder & Lightning, 7016 WIMR
- Attune V6 Map – River, 2360 Biotech
- LSR Map – 7016 WIMR
- Fortessa Map – 7016 WIMR
- MQ Map – 7016 WIMR
- ImageStream MarkII – 7016 WIMR
Use iLab to make an appointment for the following:
- Two ThermoFisher Attune Next Flow Cytometers: “Thunder” and “Lightning”
Location: 7016 WIMRThe 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)
- BD LSRII
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
Location: 7016 WIMRThe 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)
- MACSQuant10
Location: 7016 WIMRThe MACSQuant10 will be retired on June 30, 2020. Please contact the Flow Lab to schedule training and to assist with transitioning your assay to the instrument of your choice.
The MACSQuant is a 3 laser (405nm, 488nm, and 635nm), 10 parameter benchtop flow cytometer (2 scatter and 8 fluorescence). This instrument is capable of automated sampling from 5mL, 15mL, and 50mL tubes, 96-well plates, automated staining/reagent delivery, and includes an integrated MACS magnetic bead separation column for positive selection.
MQ Optical Bench Map (pdf opens in new tab)
- ImageStream
Location: 7016 WIMRThe 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)
Sorters
The facility has three high-speed sorters, 2 at the 7th floor WIMR location, and one located in 2360 Biotech. All can sort up to four separate populations simultaneously or deposit cells into multi-well plates. Operated by staff ONLY. The 2 cell sorters at WIMR are contained in custom Biological Safety Cabinets for sorting up to BSL2+ samples. The cell sorter at 2360 Biotech is contained in a BioBubble and is limited to BSL-2 samples. Note: Samples exceeding BSL2+ levels when aerosolized cannot be sorted at either location of the UWCCC Flow Cytometry Laboratory.
Sorter Maps (links open pdf in new tab):
- Aria “Jill” Map – 7016 WIMR
- Aria “Jack” Map – 7016 WIMR
- Aria “Jayne” Map – 2360 Biotech
Use iLab to make appointments on these instruments:
- BD FACSAria with Biosafety Cabinet “Jill” in 7017 WIMR
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
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” 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: 03/20/19
- 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: 02/05/19
Cell Counter: Nexcelom Cellometer
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.
Pricing
Hourly Rates |
||||
Service |
UWCCC Member |
Non-Member |
External Academic |
Off Campus |
| Attune Training/Assisted | $65.00 | $100.00 | $155.00 | $180.00 |
| Attune Self | $45.00 | $70.00 | $108.50 | $126.00 |
| MACSQuant10 Training/Assisted | $65.00 | $100.00 | $155.00 | $180.00 |
| MACSQuant10 Self | $49.00 | $75.00 | $116.25 | $135.00 |
| LSR Training/Assisted | $65.00 | $100.00 | $155.00 | $180.00 |
| LSR Self | $52.00 | $80.00 | $124.00 | $144.00 |
| Fortessa Training/Assisted | $65.00 | $100.00 | $155.00 | $180.00 |
| Fortessa Self | $52.00 | $80.00 | $124.00 | $144.00 |
| Cell Sorting (Staff)* | $94.25 | $145.00 | $224.75 | $261.00 |
| Sort Aria Self (Trained Users Only) | $75.00 | $110.00 | NA | NA |
| ImageStream Training/Assisted | $65.00 | $100.00 | $155.00 | $180.00 |
| ImageStream Self | $45.00 | $70.00 | $108.50 | $126.00 |
| Sample Prep | $65.00 | $100.00 | $155.00 | $180.00 |
| Analysis | $65.00 | $100.00 | $155.00 | $180.00 |
| Consultation | No Charge | $30.00 | $155.00 | $180.00 |
| *Cell Sorting includes 0.5 hour set up fee | ||||
Per Usage Fees |
||||
Service |
UWCCC Member |
Non-Member |
External Academic |
Off Campus |
| Lecture Series | $35.00 | $85.00 | $131.75 | $250.00 |
| Cellometer Cell Counter | Customer purchases own slides | |||