Advancing microarray assembly with acoustic dispensing technology.
In the assembly of microarrays and microarray-based chemical assays and enzymatic bioassays, most approaches use pins for contact spotting. Acoustic dispensing is a technology capable of nanoliter transfers by using acoustic energy to eject liquid sample from an open source well. Although typically used for well plate transfers, when applied to microarraying, it avoids the drawbacks of undesired physical contact with the sample; difficulty in assembling multicomponent reactions on a chip by readdressing, a rigid mode of printing that lacks patterning capabilities; and time-consuming wash steps. We demonstrated the utility of acoustic dispensing by delivering human cathepsin L in a drop-on-drop fashion into individual 50-nanoliter, prespotted reaction volumes to activate enzyme reactions at targeted positions on a microarray. We generated variable-sized spots ranging from 200 to 750 microm (and higher) and handled the transfer of fluorescent bead suspensions with increasing source well concentrations of 0.1 to 10 x 10(8) beads/mL in a linear fashion. There are no tips that can clog, and liquid dispensing CVs are generally below 5%. This platform expands the toolbox for generating analytical arrays and meets needs associated with spatially addressed assembly of multicomponent microarrays on the nanoliter scale.
Wong
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Penn Center for Molecular Discovery, Institute for Medicine and Engineering, Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Diamond
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D000162:Acoustics / Q000295:instrumentation; D056668:Cathepsin L; D002403:Cathepsins / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry; D006801:Humans; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*
D005456:Fluorescent Dyes; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C534281:CTSL protein, human; D056668:Cathepsin L; D005990:Glycerol