RE: Detecting eosinophils on a fiber optic flow cytometer

if you plot log side scatter versus green (525nm) fluorescence you can
see them as well w/o polarisation if you do not want to label them. BD
facslyse  enhances that separation.

 

Gerhard Nebe-von-Caron 

Research Scientist and Biomedical Engineer 

SPD-Spark

Swiss Precision Diagnostics 

Priory Business Park 

Bedford, MK44 3UP, UK 

Tel +44(0)1234-835474 

Fax +44(0)1234-835002 

mailto:g.nebe-von-caron@spdspark.com 

 

-----Original Message-----
From: Howard Shapiro [mailto:hms@shapirolab.com] 
Sent: 28 April 2008 23:49
To: cyto-inbox
Subject: Re: Detecting eosinophils on a fiber optic flow cytometer

 

Bill Telford wrote-

> 

> We're attempting to analyze eosinophils by analyzing polarized and 

> depolarized orthogonal light scatter, as previously described by de 

> Grooth and Terstappen, and discussed over the years on the Purdue 

> list. We're using a LSR II, and have set up two side scatter detectors


> with polarizing filters at 90 angles of polarization, using a 

> beamsplitter or a coverslip to separate the two light components. 

> We're having difficulties in resolving eosinophils this way, in 

> combination with markers for both resting and activated eos.

> 

> After reading some old Purdue posts by Howard and others, I've become 

> concerned that the fiber optics that couple our flow cell to detectors


> may not preserve light polarization, which is essential for this 

> method. When the technique was originally proposed, most cytometers 

> did not employ fiber optics for signal transmission. Does anyone have 

> any insights into whether cytometers equipped with fiber optic signal 

> transmission can be used for this technique?

> 

> You'd have to use a polarization preserving fiber in the light 

> collection path; they are typically single-mode, which would almost 

> certainly mean you'd collect less light, but if you were willing to 

> sacrifice performance on the whole trigon (you probably wouldn't be on


> an octagon), you might be able to get it to work.

 

-Howard