Neo, Size resolution, more than optical alignment, is the issue here. Using Flow Checks to optimize alignment is okay, as long as you have established criteria defining that optimal alignment. Once achieving this, using the 10 micron particles as a size marker is not very useful. I've used 1 micron and 5 micron particles to better define size ranges. I find that using the 1 micron particle allows us to better establish threshold limits and gates. Keep in mind . . . beads rarely interact with light the same as cells, so using bead "size" may mislead you re. actual cell size. MAK. Neo Sim Yee wrote: > Dear all, > > I have an enquiry with regards to the flow check paticles that are used for > assessing the optical alignment of the flow cytometer. > > In our work, we are dealing with bacteria of a size range as small as 0.5um. > The flow check we are currently using is the 10um Flow Check Fluorosphere > from Coulter. Noting the relatively large difference in size of the flow > check particles and the cells that we are analysing, does this introduce a > significant amount of error in the sense that the instrument may not be > optimally aligned fot the analysis of such small paticles? > > Thank you for your help. > > Kind Regards > > Neo Sim Yee, Lezlie > Department of Civil Engineering > National University of Singapore -- Mark A. KuKuruga, Managing Director University of Michigan Core Flow Cytometry <http://www.cancer.med.umich.edu/flow_cytometry> phone: 734-647-3216 fax: 734-936-7376 kukuru@umich.edu
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