461 ISSN 1473-7159 © 2011 Expert Reviews Ltd www.expert-reviews.com Editorial 10.1586/ERM.11.30 Application of 3D hydrogel microarrays in molecular diagnostics: advantages and limitations Expert Rev. Mol. Diagn. 11(5), 461–464 (2011) Microarrays are among state-of-the art techniques addressing the bottlenecks of efficient and highly parallel identification, measurement and analysis of significant molecules in ‘omics’-based diagnostics. According to Schrenzel et al. “microarrays consist of an orderly arrangement of probes (oligonucleotides, DNA fragments, pro- teins, sugars or lectins) attached to a solid surface” [1] . Over the past 20 years, in the bio- marker discovery, qualification and vali- dation domain, there has been a constant evolution towards the increase of sample through-put, ‘shrinkage’ of both detectors and sample volumes, and increase in sam- ple types with emphasis on biological fluids for noninvasive detection. In this complex picture, microarrays have been one of the main biological tools in the last decade [2] . Future array systems will be used for high-throughput functional annotation of gene products. The concept of using sur- face immobilization of probes is common to a broad range of applications based on molecular recognition; diagnostics repre- sents a major field in molecular recognition, but is not limited to this. Originally, chips and arrays were developed on planar sur- faces and probe fixation was achieved based on different interactions, including physi- cal (such as adsorption) and chemical (e.g., covalent or ionic), due to specific surface activation and binding of reactive groups [3] . Biological integrity of molecules triggered the need for developing hydrogels The probes are immobilized on a range of different substrates or coated slides including poly-l-lysine, nitrocellulose, hydrogel, polyacrylamide gel, agarose, aldehyde, epoxy or other polymeric coat- ings. In recent years, the use of gel matrices in slide preparation has often been cited, mostly using gels of a hydrophilic nature. Hydrogel coatings include polyacrila- mide, polyuretan, agarose, sephadex and polyethyleneglycol. Hydrogel-coated slides allow the use of multiple buffer types with- out pH modification or without the use of tertiary amine-free buffers [4] . The need for the development of hydrogel-based microarrays comes from the delicate structure of proteins that comprise biological functions in intimate relation to their complex structure. These proteins are difficult to immobilize on a solid support without harming their function. Keeping their structure intact and, consequently, their biological function is essential, thus testing them in a liquid environment is a prerequisite. The development of these tools was designed to measure complex arrays of proteins, precisely and simulta- neously for applications in comprehensive proteomics, protein networks and path- ways. Microarrays are highly involved in the validation steps of genomic discoveries and clinical biomarkers [5] . As stated, the need to develop these hydrogel microarrays came from the disadvantages of 2D acti- vated epoxy or aldehyde glass slides that immobilize proteins through covalent or electrostatic interactions. In 2D microarray KEYWORDS:฀3D-hydrogel฀•฀biomarker฀•฀biomolecule฀•฀coating฀•฀gel฀matrix฀•฀immobilization฀ •฀microarray฀•฀molecular฀diagnostics Cristiana Pistol Tanase Author for correspondence ’Victor Babes’ National Institute of Pathology, 99-101 Splaiul Independentei, Sector 5, Bucharest, Romania Tel.: +40 213 194 528 Fax: +40 213 194 528 bioch@vbabes.ro Radu Albulescu ’Victor Babes’ National Institute of Pathology, 99-101 Splaiul Independentei, Sector 5, Bucharest, Romania and National Institute for Chemical-Pharmaceutical R&D, Bucharest, Romania Monica Neagu Department of Immunology,’Victor Babes’ National Institute of Pathology, 99-101 Splaiul Independentei, Sector 5, Bucharest, Romania Authors contributed equally to this manuscript For reprint orders, please contact reprints@expert-reviews.com