FAQS
General FAQs
What are the Stem Cell Core Facility hours of operation?
How do I arrange to run samples at the core facility?
How will I be charged?
How many cells should I have and what medium should my cells be in?
What tubes should I bring my samples in?
What flourochromes can I use?
What controls do I need?
How can I store my data?
How do I cite the Stem Cell Core in a paper?
What are the Stem Cell Core Facility hours of operation?
The Stem Cell Core Facility is open from 9am – 5pm Monday through Friday.
How do I arrange to run samples at the core facility?
Prior to booking an appointment, you must first create an account by filling out a User Registration Form. You will need to provide us with contact information for your department’s financial coordinator/grant manager, billing address, email and phone number. Once you have established an account, you may book time using our online scheduler
How will I be charged?
Users are billed on a monthly basis. Please see Policies/Billing page for costs.
How many cells should I have and what medium should my cells be in?
A good starting point is 1 x 10e6 cells/ml with a minimum volume of 500ul. Fewer cells mean longer collection times, but a too concentrated solution can lead to aggregation and clogging of the sorter nozzle. For cells that are to be sorted, the starting concentration will depend on how many cells you need to recover. It is always best to estimate a lower yield rate and be sure to have an ample number of cells to start. Factors that ultimately reduce yield include electronic aborts – cells which arrive in the laser beam too close to one another, and pre and post sort cell death. A good estimate is to start with double or even triple the number of cells that you need to get back. Cells should be supplied in PBS or HBSS. All samples must be filtered through 40 um mesh before submitting them.
What tubes should I bring my samples in?
Cells should be brought in 12×75 mm round bottom tubes with snap cap (BD 352054)
What flourochromes can I use?
For information on fluorochrome excitation, emission wavelengths and spectral overlap see bdbiosciences.com
What controls do I need?
Control samples are necessary for proper instrument set up. Included should be an unstained sample (negative control) and a single stained control for each color used. Positive controls are used for compensation of flourochrome emission overlap. Please make sure that there are adequate cells (100k or more) in the control samples!
How can I store my data?
If you wish to save your data for future use, we recommend that you use a USB drive as we are not able to store data indefinitely. FlowJo software is available to download from flowjo.com for users who want to analyze their data away from the lab on their own computers.
How do I cite the Stem Cell Core in a paper?
When you publish, please acknowledge the core facility in every publication that contains data acquired in the Stem Cell Core Facility. A suggested acknowledgment is: “Flow cytometry was performed in the Stem Cell Core Facility at Children’s Hospital Boston that is supported by National Institutes of Health award NIH-P30-HD18655.
FAQs for Cell Sorting
What type of cells can I sort?
How many cells do I need and what yield should I expect?
What information do I need to provide about my samples?
What else do I need to bring?
What type of cells can I sort?
Theoretically, any cells that can be kept in a single cell suspension can be sorted. We ask that you filter your samples just prior to sorting as clumping of cells can clog the nozzle, disrupt the sort, and in some instances, necessitate the re-establishment of instrument settings.
How many cells do I need and what yield should I expect?
How many cells you need to start with depends on how many cells you would like to recover from the sort. There are many factors that affect % yield. The healthier the cells are going into the sort, the more viable they are post sort. Electronic aborts, pre and post sort cell death, cells sticking to the sample tube and low frequency of the target population are just some of the variables that can contribute to lower than expected yields. Starting with about 2x – 3x the number of desired cells will help ensure that you retrieve your targeted amount of cells.
What information do I need to provide about my samples?
The Stem Cell Core Facility is an open core facility and as such, it is imperative that we are made aware of any biohazards associated with samples that are brought in for analysis/sorting. Proper documentation helps us to prepare accordingly and we ask that you fill out a Cell Sort Form containing information about the sample source and potentially infectious agents online at our website or by delivering a completed form to the facility prior to bringing us your samples. Acceptable samples are non-primate cells (murine, zebrafish, etc.), established human cell lines and replication incompetent cells transformed with biosafety BSL1/BSL2 approved protocols. Currently, the instruments and facility cannot accommodate patient samples, infectious samples, radioactive samples or any BSL 3 material.
What else do I need to bring?
Collection tubes containing support media. 12×75 mm or eppendorf tubes are suitable, but use the smallest tube possible to collect your cells so as to lessen excessive cell loss. Bringing extra tubes and collection media is advisable. You will also need to bring a negative, unstained sample and a positive, single stained control for each color used. Please label all tubes with an alcohol-resistant marker.