Hi LuAnn, You are looking at some rather expensive options. It's always nice to get another piece of equipment, but from the information you provide I would be hesitant in recommending you purchase more equipment. I have been measuring calcium signaling through gpcr's for quite some time. We have been using the Fura Red AM/Fluo-3 AM protocol as described by Rabinovitch et.al. in Cytometry a few years ago, with great success. As you know, these dyes are excited by the argon laser and emit at ~670nm and ~530nm (approximately) respectively. In the paper, Rabinovitch describes a way to measure calcium responses with PE labeled antibody stained cells, but i've found that to be a bit tricky. it seems like you are just getting started in measuring calcium responses, there are many tricks and tips i would recommend. firstly, get a hold of as much literature as you can. i have a web page i made for the boston users group for cytometry at: "http://www.shore.net/~bugbytes/calcium.html", that will give you some of the references I found useful. So....all in all, a new laser might not be a bad idea as it could give your facility/lab added flexibility in terms of experimental design. A new machine alleviates potential scheduling issues for your users. The LSR is a wonderful machine that was meant (my perspective) to measure calcium responses with Indo-1 AM plus additional multiparametric analyses. The LSR does detect Indo-1 AM fluorescent emissions well, but it's expensive, as are your other suggestions. Ultimately the choice is yours, but there are many options. Good luck. Chris Chris Groves Millennium Pharmaceuticals Inc. 75 Sidney Street Cambridge MA 02139 617-679-7495 groves@mpi.com luann wrote: > Hi Flow-ers, > > Our flow lab is in the fortunate position of being able to add another > analyzer to our core facility. One of our researchers would like to add uv > capabilities to the lab, specifically for Ca++ measurements. At present we > are trying to determine the best method to to accomplish this. The lab > currently has an Elite sorter with gated amplifier that has an Argon and a > HeNe laser, and an XL analyzer. We are currently evaluating whether it is > preferable to buy a HeCd for the Elite and then buying a separate analyzer > ( XL or FacsCalibur), or wherther we should purchase the BD-LSR (with three > lasers). Since this is a new machine, I'd appreciate any comments from > those who have this instrument, as well as comments from others who have > faced a similar choice. I am most concerned with issues regarding > reliability, easy of use, and utility of the LSR. > > Comments may be sent directly to me at: > > luann+@pitt.edu > > Thanks all, > > LuAnn
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