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UW Health SMPH
American Family Children's Hospital
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Equipment

Introduction 

Training

Analysis Computers

Benchtop Analyzers

Sorters

Microscopes

 

Introduction

Please review Flow Cytometry - Using the Facility before making an appointment. Clicking on any of the equipment below will link you to the online scheduling system to make an appointment. The system sends email reminders automatically. Please do not email the scheduler because the staff does not respond to that email. Direct all queries to staff email addresses.

 

You MUST attend the Introduction to Flow Cytometry lecture series before signing up for any training on any cytometer. (Training on microscope is excluded.) Once you have attended the lecture series make an appointment for two hours on:


01. FACSCalibur-WIMR-Training/Assisted schedule (Tuesday 12-3pm) 
04. LSRII-WIMR-Training/Assisted schedule (Wednesday 12-3pm)

06. Fortessa-Training/Assisted schedule (Thursdays, 12-3pm)

08. MACSQuant-WIMR-Training/Assisted schedule (Thursdays, 12-3pm)

13. BD Pathway-WIMR-Training/Assisted schedule (Mondays, 12-3pm)  

 

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Training

The two-hour hands-on training is intended for those with little or no flow cytometry experience. It will teach the basics of instrument set-up and data collection. Fluorescent Beads will be used for training. Please do not bring cell samples.

 

Once the two-hour training is completed or for those who have some experience operating a flow cytometer, it will be necessary to schedule an hour of assisted time with user samples to discuss specific assay data collection. No user will be allowed to operate the cytometers independently until these training sessions are completed.

 

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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 McArdle laboratory. Computers are loaded with a variety of analysis packages including FlowJo, CellQuestPro, FCS Express, ModFit, WinList, and Image analysis, as well as standard Microsoft Office software.

There is no charge for the use of these computers. However, they must be used exclusively 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. 

 

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Benchtop Analyzers

The benchtop instruments used for this purpose are the FACSCalibur (488nm and 633nm lasers),  LSRII (488nm, 405nm, 561nm, 355nm, and 642nm lasers), Fortessa (488nm, 403nm, 561nm, 355nm, and 643nm lasers with High-Throughput Sampler for automated sampling of 96 and 384 well plates) and MACSQuant 10 (488nm, 405nm, and 633nm lasers with automated tube and 96 well plate sampling option and in-line magnetic bead separation column option). 


When signing up for appointments here, insure all fluorochromes to be used can be excited with the lasers listed. The instruments are available for self-run appointments for which no assistance is provided or for staff-assisted appointments for those who need assistance in data collection or instrument operation. Staff-assisted appointments can be used by those who have not received any instrument training but need samples analyzed, as well as by those who need assay-specific instruction.

 

Instrument Maps:

Make an appointment for:

01. FACSCalibur-WIMR-Training/Assisted schedule

02. FACSCalibur-7016 WIMR, Self schedule

All users operating cytometers after staff hours (8pm-6am) are responsible for shutting down the cytometer after use. Please contact staff with any questions.

 

03. FACSCalibur-McArdle, Self schedule


04. LSRII-WIMR, Training/Assisted schedule

 

05. LSRII-7016 WIMR, Self schedule
All users operating cytometers after staff hours (8pm-6am) are responsible for shutting down the cytometer after use. Please contact staff with any questions.

 

06. Fortessa-Training/Assisted schedule

 

07. Fortessa-Self schedule

All users operating cytometers after staff hours (8pm-6am) are responsible for shutting down the cytometer after use. Please contact staff with any questions.

 

08. MACSQuant Training/Assisted schedule

New User Training: Schedule two hours. 
Those with little flow cytometry experience: You MUST attend the Introduction to Flow Cytometry lecture series prior to signing up.

Assisted appointments: For those who need assistance operating benchtop instruments for data collection.

 

09. MACSQuant Self schedule
The MACSQuant is a benchtop cytometer is with tube and plate loaders, integrated MACS magnetic bead separation column. 488nm has four detectors, 405 nm has two detectors and 633 nm has two detectors. Trained users only.

 

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Sorters

The facility has two high-speed sorters: one located in the McArdle laboratory and one at the 7th floor WIMR location. Each sorter has some unique capabilities for the maximum applications, but they both can sort up to four separate populations simultaneously. Operated by staff ONLY. They are both housed under BSL2 conditions. Note: Samples exceeding BSL2 levels when aerosolized cannot be sorted.

 

Sorter Maps:

Make an appointment for:
10. Sorter-Aria II, BSL1 and limited BSL2

The SORP BD FACSAria is a digital system equipped with five lasers (UV, 405 nm, 488 nm, 532 nm and 640 nm). The Aria can simultaneously collect fluorescent emissions from 16 different fluorophores.  Please contact Flow Lab staff us if you have questions regarding fluorophore compatibility. 

 

11. Sorter-Aria III in BSC-BSL2

The FACS AriaIII is a cell sorter equipped with four lasers (403 nm, 488 nm, 561 nm and 637 nm). This sorter is contained in a custom Biological Safety Cabinet. Sorts containing human cells, retroviral transfections or as determined in your lab's Biological Safety Protocol MUST be sorted on this instrument.

 

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Microscopes
Please consult with the staff before scheduling an appointment here if you have not used the microscope before. 

Make an appointment for:

12. BD Pathway - 7016 WIMR, Self schedule

The Pathway is a microscope with a temperature-controlled chamber and liquid handling capabilities. This inverted fluorescent scope requires slides or 96/384 well microtiter dishes. It is equipped with two mercury arc lamps, 16 different excitation filter sets and eight different emission filter sets. It uses standard immunofluorescent probes, like Texas Red, Hoechst, and the Alexa dyes and can collect either regular or confocal images.

13. BD Pathway-7016 WIMR, Assisted 

 

14. Fluorescent Microscope-7016 WIMR schedule

The Olympus BX51 Fluorescent Microscope is capable of brightfield, darkfield and fluorescent microscopy. Attached to a CCD camera and PC for taking pictures. Self-run appointments only.

 

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FACSCaliburFirtessaFortessaMACSQuantAriaIIFACS AriaIIIBD Pathway