The SBP Flow Core will be hosting the 46th Annual Course in Cytometry this August 12-17th. Most core services will be unavailable from August 7 to August 21st, and we will be playing catch-up with our internal SBP users for a week or two after.
Come late-August we should be back to our normal lead times for appointments.
New Customers
Prior to your first visit:
If this is the first time your company/institute has used an SBP core you will need to submit a completed New Customer Form to the core.
All users must have a valid ilab account. Register for an ilab account here. It takes a few days after we receive your new customer formfor your ilab account to be activated.
After logging in to your ilab account you can request a sort, assisted analysis, or training. You will need to have a PO ready to enter into the appropriate field. If you need a quote to obtain a PO, please contact the core with information about your project.
The core is open to both SBP researchers and scientists from other institutions. We are booking 1-2 weeks out for sorts and analysis appointments.
Sort Time Calculator
I recently came across this awesome sort time calculator on the Memorial Sloan Kettering Flow Core website. It estimates how long a sort would take given the sorter settings and the number of cells you have. It’s a good reality check for whether your experiment can be done in a reasonable time. Note that these times are for ideal samples that run without clogging, or aggregation. Real sort times will probably be slightly longer.
To use the calculator, input the total number of samples, and the total number of cells (in millions) in all samples combined. So if you have 3 samples, each with 10MM cells (30 million total), enter in 3 for samples and 30 for the number of cells.
You will also need to select the nozzle size. 70um for naive primary T cells, 85um for B cells where post-sort viability is important, 100um for adherent cells.
New Customers
Prior to your first visit:
If this is the first time your company/institute has used an SBP core you will need to submit a completed New Customer Form to the core.
All users must have a valid ilab account. Register for an ilab account here. It takes a few days after we receive your new customer formfor your ilab account to be activated.
After logging in to your ilab account you can request a sort, assisted analysis, or training. You will need to have a PO ready to enter into the appropriate field. If you need a quote to obtain a PO, please contact the core with information about your project.
COVID Regulations
Current COVID regulations: visitors do not need to provide proof of vaccination. If you have had COVID recently and are now asymptomatic you must wait 10 days after first experiencing COVID symptoms or first positive COVID test before coming into the lab. If you are still symptomatic after 10 days you must wear a tight-fitting mask while on campus.
The SBP Flow Core is open to external users. We are currently booking ~1-3 weeks out.
Prior to your first visit:
If this is the first time your company/institute has used an SBP core you will need to submit a completed New Customer Form to the core.
All users must have a valid ilab account. Register for an ilab account here. It takes a few days after we receive your new customer formfor your ilab account to be activated.
After logging in to your ilab account you can request a sort, assisted analysis, or training. You will need to have a PO ready to enter into the appropriate field. If you need a quote to obtain a PO, please contact the core with information about your project.
When you visit the SBP campus, prior to entering the building each day you must:
not be experiencing any Covid symptoms
have not received a positive Covid test in the past 10 days
These are the current procedures for sorting at SBP (revised September 2022). We are updating this page with the most current information as needed. Please refer back before your appointment.
Prices – FACSAria Sort with Core Staff Operator
SBPCancer Center
Non-Profit
For-Profit
First Hour
$148
$200
$390
Each Additional Hour
$85
$115
$224
Plate Sorting + (per/hr)
$10
$15
$27
Scheduling
Our sorting services are available to researchers at both non-profit institutes and for-profit companies when we have excess capacity after scheduling SBP scientists. If your company hasn’t used SBP cores before, please see this page for instructions on setting up a new account.
Most short sort appointments can be accommodated within two weeks and longer sorts within three weeks of receiving the ilab request. State your scheduling preferences and constraints (i.e. time points, cells not ready until a certain date etc.) in the ilab request scheduling text box. We will try our best to accommodate your first choice and are most likely to be able to schedule your sort on the first try if you provide several options for your availability.
Visitors are currently allowed in the building and you are welcome to join us in the sorting lab to provide input on setting gates to obtain the population you wish to enrich. Being present during the setup phase of the sort is helpful when questions arise.
If you do not wish to join us in the sorting lab while we get set up, please provide complete written instructions (using forms linked in the next section) so that we can unambiguously understand your intentions.
Experiment Worksheets
Completed Tube and Gating Worksheets must accompany your samples if you will not be present during the sort. Bring hardcopies to your appointment. Include a reliable phone number and email on the tube worksheet so we can easily reach you if we have questions. For complicated sorts, we strongly encourage you to be present for the first hour while we set sort gates.
Illustrate your gating strategy for sort populations either with FACS data acquired previously or in cartoon form using our Gating Worksheet.
Label X and Y axes with fluorochrome and for antibody include specificity (i.e. DAPI, Actin-GFP, CD4-PE)
Label your gates as you would like us to name them (i.e Live, CD3+, Naive SORT etc.)
Indicate hierarchy with arrows from parent gate to daughter plot.
SORT Gate names on worksheet should match collection tube labels.
Labeling Sample Tubes and Collection Tubes/Plates
Samples and controls should be in 5 ml polypropylene FACS tubes (Falcon 352063) with an airtight cap. The airtight cap must be sealed prior to arrival. If you are using filter-cap tubes (Falcon 352235) you can apply some parafilm after filtration to make the lid airtight.
Label your tubes with ETOH-resistant ink or stickers before you arrive. Labeling on sample tubes must match the Tube Worksheet. We use lots of ETOH to keep our hands and surfaces clean and writing from standard Sharpie markers will rub right off. The VWR Lab Marker works well.
Collection Tubes and Plates
Label before you arrive. For collection tubes, write the sample name and the sort population name on the tube consistent with the naming convention in your tube/gating worksheets.
For collection plates, write the sample name and plate number if there are multiple plates. You can annotate the plate lid to indicate how many cells per well and which sort gates go in each group of wells. Or you can use our Sort Plate Worksheet or your own illustration that clearly shows your sorting strategy.
Checklist
Review the Sorting Checklist before you leave your lab and make sure you bring the needed supplies and materials to your sort appointment.
Proceed to the flow lab on the second floor and hand off your samples and collection tubes to core staff, or place them along with tube worksheet, and gating worksheet on the sample drop-off cart outside the sorting room.
External (non-SBP) Users
Call us from the parking lot when you arrive. (858) 646-3143. Core staff will meet you on the landing outside the lobby doors and receive your sorting samples and supplies or escort you up to the lab.
Sample Pick-up
We will email you through when your sort is done either via direct email or ilab messaging. We can also call you if that is your preference.
Sample pickup is basically the same as sample drop-off. SBP staff can pick up their samples from the drop-off cart. External users should call us from the parking lot when they arrive.
You can do a quick performance check of the Fortessa or X20 with CST beads.
Log into DiVa software,
In the browser window, look for and expand the “Shared View” section
Open the CST experiment
Look at the recorded data from prior days. Sharp peaks around 10^5 indicate good performance in all of the channels.
In the Acquisition Dashboard window, click on the NEXT TUBE button to make a new tube with today’s date.
If there is a pre-made tube of CST beads in the mini-fridge with >300ul volume remaining use that. If not make a new tube by adding 2 drops of CST beads to 1 ml of PBS.
Write “CST” today’s date, and bead lot number on the tube.
Load the CST tube on LOW and hit ACQUIRE. The peaks should look similar to the way the looked in the past. Record 2000 events
If the peaks are very broad in multiple channels there may be an air bubble in the flow cell. Remove the CST tube, keep the SIP arm to the side and press the PRIME button (next to STANDBY and RUN).
Repeat PRIME 2 more times. Then load a tube of water and run for 3 minutes.
Recheck the CST beads and see if the peaks have sharpened up.
If the peaks look reasonably sharp, it’s a good idea to run the full CST algorithm following these instructions (takes 10 minutes and need >300ul of beads with event rate >100 per second) to calibrate the laser delay settings.
Return the CST beads to the fridge. They can be reused for several days if kept in the cold and dark.
External (non-SBP) researchers with proof of vaccination can come into the core to independently operate the LSRFortessa cytometer after completing training. Use this link to request training. The Fortessa is available to non-SBP folks Monday through Friday from 10am to 2pm. Group size limit is 2 (fully vaccinated) people at a cytometer.
Sorting operated by core staff is available to scientists at SBP, and other non-profit and for-profit entities. To get in the sort scheduling queue, use this link to create an ilab sort request.
If your company/institution hasn’t previously used an SBP core, you will need to complete our New Customer Form (.xlsx) and return via email to Yoav Altman.
Each individual user will need to register for an account with iLab which is the project management software we use to schedule experiments in the core.
Getting credentials approved usually takes about a week.
Once your ilab account is active we can help you create a service request for your sort or analysis. You will receive an automated email from no-reply@ilabsolutions.com with a link to complete your service request by entering the technical details we need to schedule your experiment.
You will need to have a PO ready to enter into the ilab request.
The SBP Flow Cytometry Shared Resource is open for business. We provide cell sorting, flow cytometry analysis, and imaging flow cytometry to the greater San Diego and La Jolla science community.
We’ve adopted new procedures for the COVID age to keep you and us safe. The situation is fluid and our practices will change as needed to stay in compliance with institute policy. Please refer back to this web page for our latest procedures.
SBP Flow Cytometry 