Comparison of a pump-around, a diffusion-driven, and a shear-driven system for the hybridization of mouse lung and testis total RNA on microarrays.
In the present study, we demonstrate the benefits of a shear-driven rotating microchamber system for the enhancement of microarray hybridizations, by comparing the system with two commonly used hybridization techniques: purely diffusion-driven hybridization under coverslip and hybridization using a fully automated hybridization station, in which the sample is pumped in an oscillating manner. Starting from the same amount of DNA for the three different methods, a series of hybridization experiments using mouse lung and testis DNA is presented to demonstrate these benefits. The gain observed using the rotating microchamber is large: both in terms of analysis speed (up to tenfold increase) and in final spot intensity (up to sixfold increase). The gain is due to the combined effect of the hybridization chamber miniaturization (leading to a sample concentration increase if comparing iso-mass conditions) and the transport enhancement originating from the rotational shear-driven flow induced by the rotation of the chamber bottom wall.
Vanderhoeven
Johan
J
Department of Chemical Engineering, Vrije Universiteit Brussel, Brussels, Belgium. johan.vanderhoeven@vub.ac.be
Pappaert
Kris
K
Dutta
Binita
B
Van Hummelen
Paul
P
Desmet
Gert
G
D000818:Animals; D008168:Lung / Q000737:chemistry*; D008297:Male; D051379:Mice; D008904:Miniaturization / Q000295:instrumentation; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D012313:RNA / Q000032:analysis*; D013737:Testis / Q000737:chemistry*
D012313:RNA