Chemiluminescence detection for hybridization assays on the flow-thru chip, a three-dimensional microchannel biochip.
Chemiluminescence (CL) detection is seldom used in two-dimensional solid support microarray platforms because adequate sensitivity and spatial resolution is difficult to achieve. The three-dimensional ordered microchannels of the Flow-thru Chip increase both the sensitivity and spatial resolution required for quantitative CL measurements on microarrays. Enzyme-catalyzed CL reactions for the detection of hybridizations on microchannel glass were imaged using a CCD camera. Signal uniformity, sensitivity, and dynamic range of the detection method were determined. The relative standard deviation of signal intensities across an array of 64 spots was 8.1%. A detection limit of 250 amol of target with a linear dynamic range of 3 orders of magnitude was obtained for a 3-h assay. Similar to two-color fluorescence measurements, multiple enzyme labels were employed to demonstrate two-channel chemiluminescence. A unique method for measuring the relaxation time of a chemiluminescent species is also described.
Cheek
B J
BJ
MetriGenix Inc., Gaithersburg, Maryland 20878, USA.
Steel
A B
AB
Torres
M P
MP
Yu
Y Y
YY
Yang
H
H
D002138:Calibration; D005453:Fluorescence; D008163:Luminescent Measurements; D036103:Nanotechnology / Q000295:instrumentation*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*