Nozzle design parameters and their effects on rapid sample delivery in flow cytometry.
BACKGROUND
Rapid kinetic and high throughput flow cytometry are emerging as valuable tools in biotechnology research applications ranging from mechanistic analysis of molecular assemblies to high throughput screening. Many of these new applications have been made possible by improved sample delivery capabilities, focusing increased attention on fluidic issues associated with rapid sample delivery.


METHODS
Using basic fluidic premises, we derived a model that predicted the effect of nozzle parameters during rapid sample delivery. We tested the model using the rapid mix flow cytometer and modifications were made to the equipment to optimize performance.


RESULTS
The model predicted that shorter nozzles with wide exit orifices decrease the delay before initial particle analysis and the fluidic stabilization time. Experimental results confirmed this prediction and model-based modifications allowed analysis of particles within 55 ms or 600 ms after mixing, with or without electronic gating, respectively.


CONCLUSIONS
The model along with modifications to commercial equipment will allow rapid mix flow cytometry to analyze reactions in time frames threefold shorter than previously possible. The model allows for nozzle design predictions that should allow for analysis in the millisecond time frame. Furthermore, these findings are general for all rapid delivery applications, including high throughput flow cytometry.
Graves
Steven W
SW
National Flow Cytometry Resource, Biosciences Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, USA. graves@lanl.gov
Nolan
John P
JP
Jett
James H
JH
Martin
John C
JC
Sklar
Larry A
LA
D001331:Automation / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D005453:Fluorescence; D008432:Mathematical Computing; D008863:Microspheres; D013048:Specimen Handling