Viscosity and FCM

John Ladasky (ladasky@leland.Stanford.EDU)
Mon, 16 Jun 1997 15:29:13 -0700 (PDT)

>Date: Mon, 16 Jun 1997 08:17:33 -0400 (EDT)
>From: Matthew J Shaw <shawmj@battelle.org>
>Subject: Viscosity Effects on FCM Analysis
>
> All,
>
> Does anyone have a good feel for the effects of higher viscosity
> fluids being analyzed via flow cytometric means? For example, the
> viscosity of water is 1 centipoise, and I assume that cells that are
> within water can be reliably analyzed. I believe that the viscosity
> of blood is perhaps 3 centipoise; again, I assume that cells within
> this matrix can be reliably analyzed. However, if the matrix is say
> 10 centipoise, there may be a problem with forcing the matrix through
> the FCM nozzle. Maybe a problem doesn't exist until the matrix is 100
> centipoise? Has anyone done a study like this on any flow cytometer,
> or have a good feel for this effect? What is the upper viscosity
> limit before any problems may occur? (I realize that one could dilute
> the sample with a low-viscosity liquid until the desired viscosity is
> reached - I'd rather not have to do that!) I would appreciate any help
> on this matter.
>
> Matt Shaw

Many years ago, I performed an experiment with a FACScan that sup-
ports Matt's hypothesis that viscous fluids may not flow as readily through
the flow cytometer. My "viscous solution" was detergent-solubilized nuclei
from the Jurkat cell line, stained with propidium iodide. The stock solution
was fairly concentrated, as I recall -- perhaps 10 million cells/ml. I did
not have the means for measuring the viscosity of this solution. I tried
this at full strength and at various dilutions. I weighed the tubes on an
analytical balance, ran them on the FACScan for five minutes, and weighed
them again. I used the low flow rate, which pumps 12 microliters/minute
when the sample tube contains just plain water. Over the course of the
five minutes, I watched the acquisition event rate in order to be sure that
any drop that I might see in the *mass* flow rate (microliters/minute) was
not the result of temporary clogs in the sample injection tube. I think
that I did get such clogs at the higher cell concentrations. However, even at
the lower concentrations, there was clearly a drop in the mass of liquid
transferred, even when there were no hiccups detectable in the event rate.

Hope that helps!

- John Ladasky