Analyzer Training- We normally like to do instrument training 1 on 1 with actual samples. That way we can tailor the training specific to your assay and set up a template and instrument settings with you. We highly recommend the educational seminars prior to training.
Step 1- Contact the lab and request training (firstname.lastname@example.org)
In order to get your ID card activated for WINTROBE you need to go to the core facilities administration office located at Old Skaggs, Bldg 582 Rm 250. Terra Curley (3-2926) or Audrey Grisham (3-2928) will need your ID number and also the 6 numbers following the 2* on the back of the card.
If you need access to HCI contact Jeni Urry- email@example.com or Ali Homsombath- firstname.lastname@example.org”
If you need access to SMBB email James Marvin at email@example.com.
I will try and make this brief but there are a lot of details that need explaining. Currently, once a year Flowjo sends me a bill at the end of the billing cycle. This bill shows how many quarters a particular computer was registered with the Utah site license. If you deleted your computer or signed up half way through the year you were only billed a partial amount. Moving forward, billing will happen at the beginning of the year and registrations and authentication of the software will be handled through creating and logging into a “Flowjo Portal” account. I will be sending you an invite through my administrative tools to create your account shortly. IT IS YOUR RESPONSIBILITY TO ACTIVATED AND DELETE COMPUTERS WHEN YOU MOVE LABS OR STOP USING FLOWJO. IF YOU DONT, YOUR PI WILL GET BILLED.
The one really nice feature is that you can have multiple computers or “authorized devices” tied to your account. So you can have an office computer, laptop and home computer all registered but only pay for the equivalent of one computer. I have a feeling this might save a few labs some money. If you are a lab that doesn’t do a lot of flow you can also have a “shared computer”. Everyone in the lab would have to create a portal account to log into the shared computer but your lab would only pay for one computer. (If you think your lab would like a shared computer, please let me know).
This is going to be a great time to purge all the computers that are getting billed but are not actually getting utilized.
So here is what needs to happen.
Create Flowjo Portal account. I will send you and invite. If you don’t get an invite let me know.
Sign-in to your FlowJo Portal account through my invite and accept the group license invitation by clicking the blue banner/link.
Open FlowJo 10.5.2 or higher and follow the attached PDF instructions for FlowJo sign-in. *Delete your serial number from the serial number field, within the FlowJo license screen before signing in with your username to avoid license confusion.
After you sign-in to FlowJo with your FlowJo Portal username/password, FlowJo will communicate your computers HWA to your FlowJo Portal account. You can manage the computers that your account allows access to via your FlowJo Portal account/Authorized Devices. Each user will be able to access FlowJo on up to 4 computers of their choosing annually one computer at a time.
Shared Computers can be used by multiple users, one at a time. The computer holds the license, so any users with a FlowJo Portal account (paid or free) can access FlowJo on said Shared Computer.
Samples- Your samples can be in 5ml tubes or 15ml conical. You should aim for a concentration around 10 million/ml for cell lines or 15 million/ml for lymphocytes. If you don’t have nearly that many cells, don’t resuspend in less than 500ul. Buffer can be anything that the cells are happy in. More detailed info here
Controls- For fluorescent protein work we need an untransfected sample to set up the instrument. For multicolor experiments we need unstained in addition to single color controls.
Collection Tubes- We can collect into eppendorf, 5ml, 15ml 50ml or 96well plates. Each collection tube should have collection media already in the tube.
Extra media- In case there is a clog and a sample needs to be resorted.
Data is backed up on a server so no need to bring USB device anymore.
The most important acknowledgement for the facility and university is referencing the shared instrumentation grant for the Imagestream for any work related to your publication. Something like this would work.
“Research reported in this publication was supported by the Office Of The Director of the National Institutes of Health under Award Number S10OD026959”
Also referencing the NCI Award Number 5P30CA042014-24 is helpful.
The HSC Flow Cytometry Core Facility is a full service, state of the art lab. We offer the complete spectrum of cytometric solutions from consultation to report generation. We are currently staffed with 4 full time employees. This level of staffing allows us to accommodate training and sample prep services in an extremely timely manner. Whereas many Flow Cores simply provide access to instrumentation, our facility has a long history of supporting any level of drop off services that generally include panel design, sample prep, data acquisition and report generation.
Facilities and Instrumentation and Services: In addition to these services, we also provide instrumentation that covers virtually any cytometric assay. First of all we offer 5 cell sorters ranging from 2 laser 4 colors all the way to 5 laser and 64 detectors. These sorters can sort into 96 or 384 well plates and have nozzles sizes from 130um to 70um for very gentle sorts or high throughput sorts. The facility has 9 traditional analyzers from both Becton Dickinson and Beckman Coulter. These range from 3 laser 8 color to 5 laser 18 colors. Most of these instruments also have high throughput samplers to accommodate 96 well plate runs. The Flow Core also has 2 instruments that are not traditional flow cytometers but serve tremendous needs in the lab. First of all the lab has an Amnis Imagestream Imaging Flow Cytometer. This instrument is invaluable for any assays where the statistical reliability and multicolor potential of flow cytometry is crucial, but localization or imaging is also needed. We also have a 5 laser Cytek Aurora Spectral Analyzer with 64 detectors. This instrument is a workhorse for virtually any assay but excels when greater than 15 colors are needed in the same tube. Finally the lab has a number of ancillary equipment including Miltenyi GentleMac tissue dissociator, incubators, centrifuges, and biosafety hoods.