It is very important to have an isotype control. I have come across some investigators who have not being using isotype controls. I have seen some using only unstained cells as negative controls which is also wrong. It is like doing any other assay without a negative control. When we have negative or non-specific binding control in other assays (ELISA, RIA etc) how can you not have isotype controls in flow cytometry. It also relates to the recent discussion we have been having on this forum regarding bad data. I think investigators need to be educated more. Indresh Kaur Ph.D. Phone: 281-483-8791 -----Original Message----- From: joan Kalnitsky [mailto:jkalnits@vt.edu] Sent: Tuesday, October 30, 2001 11:38 AM To: cyto-inbox 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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