Hi all- A user is battling an issue at hand and was curious to anyone out there with any suggestions. She is trying to do flow cytometry on dog chondrocytes. The tissues are being extracted from the femoral heads of dogs. To be exact it is the articular cartilage of the subcondural bone. She then makes circular explants and is transducing them with an adenoviral vector. Before digesting them in .2%collagenase she looks at the tissues in tact to analyze the transduction efficiency based on GFP expression, then digests them. The problem is after digestion her GFP looks absolete on the cytometry analysis and lot of debris is created through this process even after filtering them though a .45u filter. Are there any suggestions on how to do this procedure on flow? Digest the tissue without disrupting GFP expression? Any thoughts are appreciated. Nicole IMCORE Laboratory Coordinator Veterinary Biosciences Ohio State University 111 Goss Lab 1925 Coffey Road Columbus OH 43210 (614)292-9716 white.1140@osu.eduReceived on Tue Nov 6 12:58:00 2007
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