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Cell Sorter

Information about the FACS in the Shapiro Science Center - Imported from https://wiki.brandeis.edu/twiki/bin/view/Bio/CellSorter

Introduction

A Fluorescence-Activated Cell Sorter (FACS) is now in place as part of the Neurobiology Microarray/FACS Core Facility at Brandeis University. This major piece of equipment uses finely tuned lasers and advanced fluidic systems for the preparative sorting of defined populations of fluorescently labeled cells. The FACS will provide new opportunities for neurobiology research at Brandeis by permitting researchers to isolate individual cell types such as neural precursor cells and specific subclasses of neurons. These cells can be used to discover the mechanisms controlling developmental processes and to obtain profiles of gene expression in specific cells in the brain.

Protocols, policies, and rates

Rates

The rates for use are as follows:

Preparatory Procedures

  • The procedures for preparing samples are as follows:
ParameterFACSAria (high-speed)
Cell Concentration (max.)1-10 million/mL
Sample Tube12x75mm polypropylene or 15mL centrifugal tube
Collection Options0.5,1.5microfuge tubes, 5,15mL tubes; 6,12,24,96 or 384 well plates
  • Minimum sample volumes are 0.5 ml.
  • Resuspend cells very well. Clumping cells will result in clogged nozzle and loss of time to clean nozzle and restart the FACS. Pipetting/ filtering cells just before they are placed on the flow cytometer is very effective.
  • Filter cells with a maximum 70um filter. Suggested filters: 70um FALCON cell strainer (35-2350) or 35um FALCON strainer cap/ test tube (35-2235) [use polypropylene tubes for sorting].
  • Bring positive and negative controls. A negative control is an unstained sample or a sample stained only with the secondary, if this is the system you are using. To make conclusions about your experiment, the proper controls are critical. If you have any questions as to what the appropriate controls are, do not hesitate to call Noreen at 6-2305.
  • If you have samples stained with more than one fluorochrome per test tube, bring in a sample stained with each fluorochrome individually. This is for compensation purposes.
  • Precoat collection tubes with BSA or serum. This will stop cells from sticking to the test tube.
  • Add 1mL of concentrated protein to the collection tube, either 1 mL of serum or 5% BSA. For sensitive cells, like neural cells, add 0.5mL of serum + 0.5 mL medium which you will use to culture cells.
  • Use appropriate size collection tubes for the number of cells you expect to collect. (i.e. a 5mL or 15mL tube). Can collect in microfuge tubes and 6 well-, 24 well-, 48- well and 96- well plates.

Policies

  • In order to use the facility, you need to have a requisition with an account number and an authorized signature on file with us. An open requisition with no set dollar amount works best for everyone. This way you do not have to bring in a separate requisition each time you use the facility, and we do not have to deal with lots of paper work.
  • Users of the Flow Cytometry Facility must give at least 24 hours notice of a canceled appointment in order to avoid being billed for the entire scheduled time and to allow others to have ample opportunity to utilize the available time. Also, billing begins from the time the appointment is scheduled to start, so please show up on time. As always, you will not be charged if another user makes use of your cancelled reservation. If you are late for your appointment and run into the next person's appointment, you must stop acquiring your samples and finish at a later time.
  • Our FACS Aria flow cytometer cannot run samples that contain radioactivity or potentially infectious agents to humans. This is because flow cytometers produce aerosols, and there is no containment system designed within them.

Computer

Log onto the computer (QUAGGA) with your UNet username and password.

Software Notes

Exporting

To export figures, e.g. to put in a presentation or thesis:

  1. select the element to be export in the worksheet (the corners of the element will show up as boxes)
  2. Choose File > Export > Worksheet Elements to save the figure as a JPEG file.

Links and References

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