Interaction of biomolecules sequentially deposited at the same location using a microcantilever-based spotter.
A microspotting tool, consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels is introduced. This spotter, called Bioplume, is able to address on active surfaces and in a time-contact controlled manner picoliter of liquid solutions, leading to arrays of 5 to 20-microm diameter spots. In this paper, this device is used for the successive addressing of liquid solutions at the same location. Prior to exploit this principle in a biological context, it is demonstrated that: (1) a simple wash in water of the microcantilevers is enough to reduce by >96% the cross-contamination between the successive spotted solutions, and (2) the spatial resolution of the Bioplume spotter is high enough to deposit biomolecules at the same location. The methodology is validated through the immobilization of a 35mer oligonucleotide probe on an activated glass slide, showing specific hybridization only with the complementary strand spotted on top of the probe using the same microcantilevers. Similarly, this methodology is also used for the interaction of a protein with its antibody. Finally, a specifically developed external microfluidics cartridge is utilized to allow parallel deposition of three different biomolecules in a single run.
Berthet-Duroure
Nathalie
N
UMR 5504, UMR 792, Ingénierie des Systèmes Biologiques et des Procédés & Plate-forme Biopuces, CNRS, INRA, INSA, 135, avenue de Rangueil, 31077 Toulouse, France.
Leïchlé
Thierry
T
Pourciel
Jean-Bernard
JB
Martin
Cristina
C
Bausells
Joan
J
Lora-Tamayo
Emilio
E
Perez-Murano
Francesc
F
François
Jean M
JM
Trévisiol
Emmanuelle
E
Nicu
Liviu
L
D000447:Aldehydes / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry / Q000378:metabolism*; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D015342:DNA Probes / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry; D019085:Fluorescent Antibody Technique, Direct; D005898:Glass / Q000737:chemistry; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D015203:Reproducibility of Results; D012825:Silicon / Q000737:chemistry; D012996:Solutions / Q000737:chemistry
D000447:Aldehydes; D000906:Antibodies; D020099:Coated Materials, Biocompatible; D015342:DNA Probes; D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D011506:Proteins; D012996:Solutions; D004247:DNA; D012825:Silicon