ProPlate® Chambers Minimize Microarray Variation

IN Featured Research, ONCYTE nitrocellulose film slides, Product Applications, ProPlate, Seals and chambers, SuperNOVA, Uncategorized
logo gracebiolabsMicroarray results are susceptible to variation introduced from multiple sources during the course of assay methodologies employed with this technology. A commonly overlooked parameter which may significantly impact results is the choice of chamber/vessel for performing microarray incubations and washes. Chambers which allow for active mixing can significantly improve microarray data. Grace Bio-Labs offers ProPlate® incubation chambers which allow for continuous mixing during microarray assays and hybridizations – significantly decreasing variability and increasing sensitivity while minimizing reagent requirements. The use of these chambers significantly reduces microarray variability (CVs ~ 10%) compared to conventional methods (CVs ~ 35%) and allows researchers to interpret their data with the utmost confidence.
Figure 1. Improved Post-Assay Spot Morphology and Decreased Spot Variation.

 

 

Spot_Morphology Improved Post-Assay Spot Morphology and Decreased Spot Variation.


Figure 1.
3-D representations and typical uniformities from microarray spots obtained after (A) incubations performed in a ProPlate® Chamber allowing for active mixing versus (B) incubations performed under a glass coverslip providing no mixing. Incubations were performed for 1 hour with anti-IgG-TRITC and -Cy5 at room temperature.
Figure 2. Reduced Microarray Variation with ProPlate® Chambers.

 

Reduced_Microarray Reduced Microarray Variation with ProPlate® Chambers.

 

Figure 2. Data show the (i) Intra-Spot variation (pixel-to-pixel), (ii) Intra-Slide variation (spot-to-spot, 10 mm between spot groups), and (iii) Inter-Slide variation (between spots from 4 arrays). All data were obtained from spots deposited on ONCYTE SuperNOVA porous nitrocellulose slides at protein concentration of 1 mg/ml (within the linear dynamic range of binding and detection). Fluorescence data are normalized and background-subtracted, collected at 532 nm from spotted goat IgG assayed with rabbit anti-goat IgG-TRITC. Assay volume under the cover slip was 50 µl (1:25,000 Dilution) and in the ProPlate was 300 µl (1:150,000 Dilution). Intra-Spot CV data are pixel variation per spot, N = 64 spot replicates (over 4 microarrays). Intra-Slide CV data are spot variation per array, N = 4 microarrays (mean of N = 16 spot replicates/array). Inter-slide data were obtained from 4 separate microarray slides.