Getting Started:

• Is this project at a biosafety level higher than BSL-2? We do not support any projects that are higher than a BSL2 category by biosafety.
• Does this project involve research animals?  If so, both a copy of your referencing IACUC (Institutional Animal Care & Use Committee approved) project and its approval will be needed before you use the instruments.

Accounts set up and instructions

Before you are able to use the Core equipment, you must complete a few basic steps.  The first of these is to set up an iLab (CrossLab) account.  This is the system used for all ICBR resources.  The registration process can be started by navigating your internet browser to http://www.biotech.ufl.edu/become-a-customer/. Here you will see information about how to become an ICBR customer, as well as instructions and links to the Registration portal at https://ufl.corefacilities.org/.

Once you have established an iLab account, you will be able to view all instruments’ schedules that you have access for or are trained to use.

All instrument booking is now conducted by the user ONLY. This includes booking any training necessary prior to unrestricted solo operation. Consultation with ICBR staff is encouraged so that we can provide any assistance needed during the time booked.

Compliance and IACUC help

All projects conducted within Cytometry must be fully compliant with the descriptions given in either EH & S registration forms or IACUC project approvals.  Help is available through the Core to assist in completing IACUC project submissions, either through direct consult or via standard language used for animal use in the Core environment.  Doug Smith and Angel Sampson specialize in this aspect and can help you move your project forward.

A generalized version of IACUC acceptable language for the in vivo imaging machines is available here: CTAC_IACUC_verbage

Facilities access help

Facilities access to the 5th floor, containing the vivarium, is controlled by the main MBI front office.  They will coordinate the 5th floor facility access form and accept once completed.  This is an online form and is entitled: “MBI Restricted Access Request Form“.  Once approved, they coordinate with Animal Care Services staff to set up a familiarization tour to compete facility specific training.

Using ICBR Resources: Signing In and Out

There are two main steps that everyone needs to perform when utilizing any equipment. You must perform both steps for the iLab system to work correctly.

1) Schedule a timeslot in the iLab calendar.

 Log into the “Self-Service Timer”.

If you wish to use one of the Cytometry systems, you must book a timeslot on the system calendar in iLab (CrossLab). Yes, even if you are only going to use the equipment for a couple of minutes; you must book a time slot on the online calendar. Go to this link to enter iLab and access the equipment calendars: https://ufl.corefacilities.org/account/login

If you have a valid iLab account, but you cannot access the equipment calendar, it may mean that you have not been granted access permission in the iLab system.

After you have booked your time slot online and you arrive at the ICBR facility, you must login to the Self-Service Timer (iPad) before you begin using the equipment. Once you have finished, return to the Self-Service Timer and log yourself out. (Operation of the Self-Service Timer is detailed below.)

Sign In

ICBR uses iLab (CrossLab) exclusively for all equipment and workstation reservations.  ALL users must therefore have an iLab (CrossLab) account in order to access the equipment in our facility.  The sign in procedure activates a timer to record the duration of your use for billing purposes.

When users come to ICBR to use their previously reserved item(s), they must start and stop a timer within the iLab platform to log the time of use.

iLab Sign In/Out System Operation:

To start the timer on your instrument session, you will need to logon using your Gatorlink credentials.

Sign in at the prompt using your Gatorlink credentials.

Windows will prepare a user session in your name.  Wait until it has finished this operation.

The computer should automatically establish a web link connection to the ICBR Self Service Application portal.  This provides links for starting the timer, entitled “Log In to Self Service Application”, or can also be used to establish a future reservation (see top right, “Login to CrossLab (iLab)”).

Should the automatic connection not autoload, the standard desktop will be seen.  In this case, look to the left side of the screen and double click the icon for the Self Service Timer.

Click on the “Log In to Self Service Application” button.

You will connect with the university’s Authentication system.  Enter in your Gatorlink ID and password.

You will be directed to ICBR’s Self-Service Check In page, where all the different ICBR cores has a presence.  Select the yellow Cytometry Core (Ct) at the bottom.

Now you will be able to see a list of all the instruments.  Scroll down and find the instrument that you wish to start a timer for, then hit its “Select” button.

Once inside the page for the selected instrument, click on “Start Time” to start the timer.  The timer will start to advance.

When you have finished your use of the instrument that you scheduled, return and repeat this process to “Stop Timer”.

Once you have stopped the timer, click on the “X” in the corner of the page to exit the application.

To close out correctly, you MUST end your Windows session.  Go to the Windows button in the bottom left of the screen and click on it.  A sidebar will be visible, and you should click on the top most control.

This will change the window so that it displays the user’s Gatorlink name.  Click on this to pop up a control box, then click on “Sign out”

You have now signed off, and the resting home screen should be seen.

To STOP the timer, repeat these steps and press the Stop button on the page with the selected instrument.  Then continue as described to logout and end your Windows session.

iLab Account Registration and Manual

Biowaste Handing, Control & Responsibilities

Management of Animal Diet Autofluorescence

Recognition of Components of the Vaporizer

Facsdivav6_lsr_quickrefguide

FCS_Express_information_email

Fluorofinder (light)

Flow Cytometry Basics Guide

Minimum Information About a Flow Cytometry Experiment (MIFlowCyt)

University of Florida Sorting Contract

Titrating Antibodies on Human PBMC

Sorting Buffer Recipe

Distributions of Autofluorescence After Compensation: Be Panglossian, Fret Not

Spectral Compensation for Flow Cytometry: Visualization Artifacts, Limitations, and Caveats

MiSet RFC Standards: Defining a Universal Minimum Set of Standards Required for Reproducibility and Rigor in Research Flow Cytometry Experiments

Rigor and Reproducibility in Flow (Tim Bushnell)

Reproducibility in Flow Cytometry through Standardization

NIH Enhancing Reproducibility Guidelines

Scientific Rigor and Reproducibility (Kemp, 2016)

Rigor and Reproducibility of Cytometry Practices for Immuno-Oncology (Laskowski 2020)

Flow Repository (Spidlen 2012)

In Vitro Imaging

Policies for Self Service Users

The Cytometry core provides a variety of tools and expertise for cellular measurements. The laboratory incorporates numerous flow cytometers, from simple entry-level devices (FACS Calibur or CyFlow-C6, BD Biosciences, San Jose, CA) to high-end 5-laser, 16-parameter instruments to analyze (LSR-II, BD Biosciences) or sort cells (Aria-II, BD Biosciences). In addition to flow cytometers, the laboratory offers a laser confocal scanning microscope (TCS-SP5, Leica Microsystems, Exton, PA). The microscope is well equipped with a variety of excitation lines from near UV to red and can collect 3 to 4 emission colors (depending on dye combinations), with viable cell time lapse and extensive computed parameter capabilities, including FRET, FRAP, image stitching, and 3D reconstruction. The laboratory offers two sites: a main site at the Cancer/Genetics Research Complex, and a satellite laboratory at the McKnight Brain Institute. Staff members are available for consultation in planning cytometric assays and carrying out the analytical portion of the assay, including analysis of the resulting data. Samples must be prepared for cytometry in the investigator’s laboratory, although if needed, some preparation capabilities exist at the Cancer/Genetics site. Training in analytical instrument operation for Investigator’s staff is also available.

In an effort to make acknowledgments easier than ever, UF | ICBR now offers Research Resource Identifiers (RRIDs)– persistent and unique identifiers for referencing a research resource. Copy and paste the appropriate core RRID (below) for citation. Proper acknowledgment of UF | ICBR in publications and awards validates our role in driving research forward and provides a platform to improve our services and instrumentation.

If you would prefer copy to include in your work:

“The data presented herein were obtained at the UF | ICBR Cytometry core, which is a UF Research core research facility at the University of Florida. We thank the UF | ICBR Cytometry core, University of Florida for the use of the [instrument name here], which provided [service].”

Authorship credit should be given if members of the UF | ICBR staff have provided services or expertise that is critical to a manuscript. Although other circumstances may warrant authorship, authors are those that meet the items below:

• Agrees to be accountable for accuracy and/or integrity of the work and expertise. (In all our work the person executing the work takes the fullest degree of responsibility for accuracy and integrity.)
• Provides substantial contributions to the experimental design, acquisition, and analysis/interpretation of the work.
• Provides input on drafting of the work (often figures) and approves of the work to be submitted/published. (If a member of the facility has made substantial contributions it is expected that s/he will be given the opportunity to review drafts of manuscripts for accuracy/fidelity and should indicate agreement before a draft is moved forward to publication).