title: Development of a high-throughput Candida albicans biofilm chip.
abstract: We have developed a high-density microarray platform consisting of nano-biofilms of Candida albicans. A robotic microarrayer was used to print yeast cells of C. albicans encapsulated in a collagen matrix at a volume as low as 50 nL onto surface-modified microscope slides. Upon incubation, the cells grow into fully formed "nano-biofilms". The morphological and architectural complexity of these biofilms were evaluated by scanning electron and confocal scanning laser microscopy. The extent of biofilm formation was determined using a microarray scanner from changes in fluorescence intensities due to FUN 1 metabolic processing. This staining technique was also adapted for antifungal susceptibility testing, which demonstrated that, similar to regular biofilms, cells within the on-chip biofilms displayed elevated levels of resistance against antifungal agents (fluconazole and amphotericin B). Thus, results from structural analyses and antifungal susceptibility testing indicated that despite miniaturization, these biofilms display the typical phenotypic properties associated with the biofilm mode of growth. In its final format, the C. albicans biofilm chip (CaBChip) is composed of 768 equivalent and spatially distinct nano-biofilms on a single slide; multiple chips can be printed and processed simultaneously. Compared to current methods for the formation of microbial biofilms, namely the 96-well microtiter plate model, this fungal biofilm chip has advantages in terms of miniaturization and automation, which combine to cut reagent use and analysis time, minimize labor intensive steps, and dramatically reduce assay costs. Such a chip should accelerate the antifungal drug discovery process by enabling rapid, convenient and inexpensive screening of hundreds-to-thousands of compounds simultaneously.
authors.affiliation: Department of Biomedical Engineering, The University of Texas at San Antonio, San Antonio, Texas, United States of America.
authors.forename: Anand
authors.initials: A
authors.lastname: Srinivasan
authors.forename: Priya
authors.initials: P
authors.lastname: Uppuluri
authors.forename: Jose
authors.initials: J
authors.lastname: Lopez-Ribot
authors.forename: Anand K
authors.initials: AK
authors.lastname: Ramasubramanian
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D002176:Candida albicans / Q000187:drug effects* / Q000254:growth & development* / Q000648:ultrastructure; D015725:Fluconazole / Q000494:pharmacology; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning
chemical_list: D000935:Antifungal Agents; D000666:Amphotericin B; D015725:Fluconazole