The need for controls depends on the specific question being asked. A significant amount of phenotyping is to obtain the percentage of cells in specific populations. With spleen cells, there are always cells negative for the markers being stained that serve as a very nice internal control. More controls are certainly needed when setting up a new assay but with experience and decent antibodies, the negative cells are an adequate control for routine research use. If you are trying to do quantitation of expression level, then additional controls would certainly be indicated. Just my $0.02. -Jim Weaver ************************************************* * * * James L. Weaver Ph.D. * * Division of Applied Pharmacology Research * * Office of Testing & Research * * CDER MOD-1, FDA * * 8301 Muirkirk Rd, Laurel MD 20708 * * * * Phone: 301-827-8237 * * Fax: 301-594-3037 * * Email:WEAVER@CDER.FDA.GOV * * * ************************************************* * -----Original Message----- * From: joan Kalnitsky [mailto:jkalnits@vt.edu] * Sent: Tuesday, October 30, 2001 12:38 PM * To: Cytometry Mailing List * Subject: Input on class experience * * * * I have a client who is teaching an undergraduate * immunology class this * fall. He approached me about including Flow Cytometry. I * was thrilled to * help and introduce a new batch of potential scientists to * Flow Cytometry. * Week number one the class came to the lab to get an overview of Flow * Cytometry. This week they were doing an experiment which * involved single * and double staining of spleen and thymocytes. When the * samples arrived I * was pretty surprised. My client, their instructor, had not done any * negative controls of any kind. No Igg controls, no * unstained cells, * nothing. When I approached him about it he did not seem to * think it was a * problem, after all he has done this staining for years. * I am curious what this listserve has to think about * this. Am I being over * vigil about the need to include negative controls in the * education process. * As far as I am concerned, there is no way to validate this * experiment to * the students. Flow cytometery is what I spend my working * day on and it * irks me to no end to see it presented incorrectly. Am I correct to * consider this presentation of Flow incorrect? Am I getting * caught up on * philosophical issues and missing the point here? * The lab is coming back again next week to learn about * apoptosis. I don't * want to open a can of worms if it isn't warranted. * Thanks in advance for all feedback. * Joan K * * * * Flow Cytometry Lab Supervisor * VMRCVM * (540) 231-4115 * FAX 540-231-7367 * jkalnits@vt.edu * * "It is better to serve than to receive." * B. Borg *
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