Perhaps fixation with 1% paraformaldehyde will help. For best results, make a 5% stock solution of paraformaldehyde by dissolving paraformaldehyde powder in PBS. Heat the solution on a hot plate under a fume hood since this produces harmful vapor. Bring to a boil. After a short while, the powder will go into solution. Turn off the heat and let it stir for about 10 to 15 minutes to completely dissolve. Filter sterilize the solution and keep it in the refrigerator. Add 5% paraformaldehyde solution to the cell mixture to a final concentration of 1%. Fix the cells for 10 to 15 minutes at room temperature and you can keep the fixed cells in cold for a day or two. This will fix the cells without making holes unlike formaldehyde or glutaraldehyde. Hope this works. SJ S. Jayaraman, Ph.D. Associate Professor of Surgery University of Illinois at Chicago 909 South Wolcott Avenue-704E MSB-M/C 790 Chicago, IL 60612 Phone: 312-355-5133 Fax: 312-355-1497 On Wed, January 2, 2008 12:59 pm, MODEL, MICHAEL wrote: > Dear List: > > > > I am trying to do something to prevent penetration of a small (mw under > 1,000) hydrophilic molecule into chemically fixed cells. The cells can > be fixed with formaldehyde or glutaraldehyde for example, but that makes > their membranes leaky, so I am looking for a way to either fix them > without opening large holes or to somehow plug the holes afterwards. > Does anyone have any experience that might suggest a possible approach? > > > > Thanks in advance and happy New Year. > > > > Michael Model, Ph.D. > > Confocal Microscopy Core > > Dpt. Biological Sciences > > Kent State University > > Kent, OH 44242 > > tel. 330-672-2874 > > > > --Received on Fri Jan 4 12:18:01 2008
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