Flow Cytometry Core Facility

Cytometry Protocols

In this section we have placed a number of protocols and reagent recipes to help users get the most from their interactions with the facility.  Choose a geneal category at the left to see the protocols we have made available.

All of these protocols and reagents have been used successfully for flow cytometry related experiments, and the bulk of them are directly out of Dr. Perry's personal protocol notebook.

Biosafety Protocols

Biosafey Sheets to Download

Policy on Biosafety and Infectious Agents

During both analysis and sorting the FACSAria flow cytometer makes up to 90,000 droplets per second.  When running unfixed cells, many of these droplets may contain viable cells.  While this stream of droplets is routinely caught in a waste receptacle and combined with at least 10% bleach to render the sample harmless, during an instrument failure (such as a partial clog of the nozzle) the potential exists for the production of large amounts of secondary aerosols.  As samples may contain viable cells which may be tumorigenic, contain pathogens and/or toxic/carcinogenic dyes, we all need to be concerned with aerosol containment for the safety of both the facility personnel and the facility users.  In order to understand and minimize the health risks to facility personnel and users the following policies have been initiated:

  1. To reduce the infectious risk to the instrument operator and the investigator, we require that all samples must be fixed prior to analysis unless prior consent has been obtained from the facility.

  2. While we do not at this time require HIV, EBV or Hepatitis testing of every sample, we do require that investigators inform the facility if samples are known to be positive.

  3. In an effort to identify risks associated with each set of samples, investigators must complete the following paperwork:

    a.  An approved “Project Biosafety Questionnaire” must be on file in the cytometry core before work on a project is started. This questionnaire must be filled out by the laboratory director (PI) and includes details about each project (i.e. a grant, project or set of experiments). This questionnaire exists in two formats:

    i.     General Project Biosafety Questionnaire – a two page questionnaire giving details on how the investigator intends to use the facility and identifying potential biosafety risks.  This form should be adequate for the majority of users not doing work with human cells or unfixed samples.

    ii.     Detailed Project Biosafety Questionnaire – a four page questionnaire designed to go into more detail regarding potential biosafety issues.  This form is appropriate to use if your project uses human cells, live cells, or genetically altered cells.

    b.  A  Daily Cytometry Sample Sheet must accompany each set of samples brought to the flow cytometry core facility.  This sheet identifies the PI and investigator associated with the samples, and gives us a contact number if we have questions while running the samples.  It also asks you to identify the sample source, whether they have been fixed, whether they have been treated with pharmacological agents, and whether they might be infectious.

We recognize that investigators are already being asked to complete a lot of paperwork, so we have tried to make this as painless as possible.  However it is clear that biosafety risks exist in flow cytometry core facilities, and we feel it is in the best interest of everyone involved to identify, minimize and contain these risks.

(last updated: November 2010)

Cell Preparation Protocols



Information on Mice

The Mouse In Biomedical Research (2nd Edition)

  • Volume 1: History, Wild Mice and Genetics
  • Volume 2: Diseases
  • Volume 3: Normal Biology, Husbandry and Models
  • Volume 4: Immunology

Information on mouse anatomy, hematology, surgery and tissue collection can be found in Volume 3. 

Tissue Culture Protocols

Phenotyping Protocols

Cell Cycle Protocols

Viability and Apoptosis Protocols

Platelet Protocols

Immunohistochemistry Protocols