Improvement of yeast biochip sensitivity using multilayer inorganic sol-gel substrates.
Today, microarray fluorescence detection is still limited because a great proportion of hybrids remain undetectable. In this paper we describe sol-gel optical multilayers (stacks of low- and high-index layers) deposited on glass slides which increase the fluorescence of DNA microarrays and favour the detection of fluorescent targets. An alternative to the expensive and time-consuming physical vapour deposition technology is proposed. It is a low-cost sol-gel coating of glass slides, each layer being made by "dipping" (alternatively in SiO2 or TiO2 solutions), "draining and drying". After the selection of the best surface layer of the substrates, the multilayer mirrors modelled for one (Cy3) or two (Cy3 and Cy5) fluorophores are spotted with a series of Yeast probes and compared to similar microarrays on standard glass slides through hybridisation experiments. The fluorescence images of the mirrors show increased signals for all the probes. The enhancement factors determined for Cy3 and for Cy3/Cy5 mirrors (10-12 and 4-5, respectively) are consistent with the initial modelling. This allows the assessment of the basal expression levels of Yeast low-expressed genes. Moreover, these substrates show a noticeable increase in sensitivity for induction/repression ratio measurements in differential gene expression experiments. So, they could be considered as promising tools for the analysis of small biological samples.
Fouqué
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B
CEA, DSV/DRDC/Biopuces, Grenoble F-38054, France. brigitte-s.fouque@cea.fr
Brachet
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AG
Gétin
S
S
Pégon
P
P
Obeïd
P
P
Delapierre
G
G
Chatelain
F
F
D005782:Gels; D020411:Oligonucleotide Array Sequence Analysis; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide; D014025:Titanium
D005782:Gels; C009495:titanium dioxide; D012822:Silicon Dioxide; D014025:Titanium