Arazdordi, Thank you for your reply. Your advice is good. Ethidium bromide, as well as PI, binds DNA and can be excited with a 488 Argon laser, it should stain the parasite nuclei, but I will have to test it and find out the fixation protocol myself. I'm hoping someone has a complete recipe. Thank you again. Gabriel >How about direct staining with ethidium bromide and enumerating with >flow cytometry!! > > >-----Original Message----- >From: Gabriel Alespeiti [mailto:gabriel_alespeiti@nybc.org] >Sent: Thursday, July 25, 2002 3:13 PM >To: Cytometry Mailing List >Subject: Re: anti-malarial antibodies > >Dear colleagues, > >I have read the replies on anti-malarial antibodies and I hope to >get some help from the experts in this field. A researcher in the >Institute is conducting parasitaemia studies of Plasmodium >falciparum infected erythrocytes using Giemsa stained microscopy, a >tedious, time consuming and technically imprecise method. We want to >replace the microscopic counts with a flow cytometry method. >Peripheral blood will be collected in Brazil and will need to be >fixed before shipping. A FACScalibur is available for this work. We >would like to analyze 25 samples a day, the expected parasitemia >level is between 0.05% and 8%. >I tried formaldehyde fixation followed by propidium iodide staining >and even though there is a direct correlation between cytometric and >microscopic determination, the manual counts are significantly >higher than the flow counts. >What I'm looking for is a simple, proven protocol for daily use that >gives directly the % of parasitaemia by flow. In our case, >determination of erythrocyte or parasite antigens; even though >useful, is not necessary. What is important for us is to be able to >detect low infection levels. >Does anybody know, what is the lowest level of infection that can be >detected by flow? >Please post the reply on this site, so other people can benefit from >it. Thank you very much. > > > > > >-- >************************************** >Gabriel E. Alespeiti >Supervisor, Flow Cytometry >Lindsley F. Kimball Research Institute >New York Blood Center >310 E.67th St. Room 2-16E >New York, NY 10021 > >Phone: (212) 570-3346 >Fax: (212) 570-3307 >E-mail:gabriel_alespeiti@nybc.org >**************************************
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