Re: Phagocytosis by Flow
Jeffrey A. Clapper (clap@iastate.edu)
Tue, 25 Jun 1996 12:11:02 -0500
>From: "JOAN CUNNICK" <jcunnick@iastate.edu>
>Organization: Iowa State University
>To: "Jeffrey A. Clapper" <clap@iastate.edu>
>Date: Tue, 25 Jun 1996 10:46:22 GMT-600
>Subject: Re: Phagocytosis by Flow
>Priority: normal
>
>Jeff: please pass this information on to others.
>
>Phagocytosis on the Flow cytometer can easily be performed by using a
>Phagotest Kit from Orpegen in Germany (Med.-Molekularbiologische
>Forschungsgesellschaft, mbH, Czerny-Ring 22, 6900 Heidelberg, Tel
>06221/9105-0, Fax 06221/910510
>
>I have used this kit equally well for human, swine, and rat blood
>samples. It supplies FITC-conjugated E.coli that are opsonized along
>with a quench solution (a blue reagent-trypan blue?) to quench the
>FITC of bacteria binding to the outside of phagocytes. Also included
>are a lysing solution to get rid of RBC when using blood samples, a
>wash/fixative, and PI solution to distinguish prokaryotes and
>eukaryotes and eukaryote aggregates. The major problem I have had
>with the kit is the expense of purchasing it in Duestch marcs, and
>paying for shipping and customs charges to get into the USA. I do not
>believe that other companys in the US sell this kit or market it.
>
>Conversely, there is a protocol in Current Protocols in Immunology
>for the FITC conjugation of bacteria. This protocol uses a
>fluorescent microscope. It should also work with a flow cytometer if
>the phagocytes are in a single cell suspension.
>
>Joan E. Cunnick; Microbiology, Immunology, and Preventive
>Medicine (MIPM); 207 Science I; Iowa State University; Ames, IA 50011
>phone:515-294-2070 FAX:515-294-6019 e-mail:jcunnick@iastate.edu
>
>From here to there, from there to here, funny things are everywhere!
>
Jeff Clapper
Cell and Hybridoma Facility
Iowa State Univeristy
515-294-8504