Today is Friday, Feb. 21, 2020

Department of Environmental Health

Center for Environmental Genetics

 

Flow Cytometry

Shared equipment and services: The CEG owns and manages equipment including flow cytometry, confocal microscopy, versatile imaging systems‎, and single cell sequencing. Access to equipment and services is free of charge for CEG members. Access requires pre-schedule and sign-up (registration link under development). See the appropriate contact persons listed on these pages for special requests/questions regarding each service or type of equipment. 

[The flow cytometer and confocal microscope are no longer eligible for ITS subsidy unless their usage involves new technology development. Please contact ITS Core director Ying Xia, Ph.D., if you have any questions about this change in ITS matching funds policy.]

Flow cytometer: The Luminex (previously Millipore) provides 3 lasers (violet, blue, and red) and 10 fluorescence channels for the detection of fluorescence-labeled cells and particles. This flow cytometer uses microcapillary to allow direct sampling by aspiration and eliminates the need of sheath fluid. The automated sampler for a 96-well plate and 10 sample tubes enables walkaway sample handling for higher throughput assays. A rotating and variable-speed mixing paddle is used between each well to help cells stay in suspension during sampling. These features provide advantages and cause errors meanwhile in sample measurement.

Sample format Samples should be prepared with 96-well sample plates (U-bottom preferred) or 1.5ml sample tubes. The cell suspension should be maintained at ~200 cell/ml and ~200 ul/well for the 96-well plate format.

Data Storage A project drive, called “Document-Flow users”, has been set up for temporary storage. If the computer space is limited, the stored data will be removed after 6-month from initial acquisition.

Data Analysis Data can be analyzed with the software GuavaSoft inCyte or Guava Suite provided by the manufacturer. Data can also be analyzed with FlowJo (group license is available from CCHMC flow cytometry core), FCS Express, and ModFit LT, which are also available for data analysis free of charge to CCHMC flow core users in R5503. See "Software" for more information.

Users are encouraged to understand the basic knowledge of flow cytometry in experimental design, antibody staining, compensation, and data analyses together with unique setting for this easyCyte unit to ensure success. These resources include the instrument website of Guava easyCyte 12-channel high throughput (12HT) flow cytometer, flow cytometry resources, on campus lectures provided by Ohio River Valley Cytometry Association (ORVCA), and related seminars from CEG.

Contact:  Shouxiong Huang (Kettering Rm 248, Rm 251), Shouxiong.huang@uc.edu; Manju Sharma (Kettering Rm 248, Rm 276) SHARMAMU@ucmail.uc.edu

Location: The instrument is located in Rm 253 Kettering

Sign-up/reservation form (link under development): Required for each use. Usage slots of more than 4 hours must be justified and pre-approved.

Training mainly involves basic instrument and software manipulation to allow new users to start sample measurement independently. At least one-hour instrument demonstration and one-hour guided hands-on sample running are required prior to independent usage. Users can request more training hours if they are unable to fully perform the assays independently.

Trained users can reserve and access the instrument from 8:00 am to 5:00 pm, Monday through Friday. Usage may be available after working hours at a case-by-case scheduling base. Users are responsible for the correct manipulation of the instrument to achieve results and avoid damages.

Staff-operated assays A staff is available to run pre-prepared samples. This service is currently offered at restricted time. Currently, the service is open every week for two-three hours slots on Monday and Wednesday afternoons.

Consultation Users are encouraged to make appointments with Dr. Huang to discuss experimental design, pitfalls, and data analyses.analyses. Contact: Shouxiong Huang, PhD

 

Research supported by the Center for Environmental Genetics should acknowledge NIEHS Award P30 ES006096