Cell analysis system based on compact disk technology.
BACKGROUND
A cell analysis system was developed to enumerate and differentiate magnetically aligned cells selected from whole blood. The cellular information extracted is similar to the readout of musical information from a compact disk (CD). Here we describe the optical design and data processing of the system. The performance of the system is demonstrated using fluorescent-labeled cells and beads. Materials and Methods System performance was demonstrated with 6-microm polystyrene beads labeled with magnetic nanoparticles and allophycocyanin (APC) and immunomagnetically aligned leukocytes, fluorescently labeled with Oxazine750 and CD4-APC, CD8-Cy5.5, and CD14-APC/Cy7 in whole blood.


RESULTS
The sensitivity of the system was demonstrated using APC-labeled beads. With this system, beads containing 333 APC molecules could easily be resolved from the background. This level of sensitivity was not achievable with a commercial flow cytometer. A maximum of 20,000 immunomagnetically labeled cells could be aligned and analyzed in between 0.6 m of Ni lines, distributed over a surface area of 18 mm(2) and extracted from a blood volume that depended on the height of the chamber. The utility of the system was demonstrated by performing a three-color CD4-CD8-CD14 assay.


CONCLUSIONS
We built a cell analysis system based on immunomagnetic cell selection and alignment and analysis of fluorescent signals employing CD-technology that is as good or better than current commercial analyzers. The cell analysis can be performed in whole blood or any other type of cell suspension without extensive sample preparation.
Tibbe
Arjan G J
AG
Biophysical Techniques Group, Faculty of Applied Physics, Twente University, PO Box 217, 7500 AE Enschede, The Netherlands. A.G.J.Tibbe@tn.utwente.nl
de Grooth
Bart G
BG
Greve
Jan
J
Rao
Chandra
C
Dolan
Gerald J
GJ
Terstappen
Leon W M M
LW
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001772:Blood Cell Count / Q000295:instrumentation* / Q000379:methods* / Q000592:standards; D001773:Blood Cells / Q000166:cytology*; D016250:Compact Disks / Q000592:standards / Q000639:trends*; D005246:Feedback; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D018189:Immunomagnetic Separation / Q000295:instrumentation* / Q000379:methods*; D008280:Magnetics / Q000295:instrumentation*; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted / Q000295:instrumentation; D012985:Software Design
D000954:Antigens, Surface