nBioChip, a Lab-on-a-Chip Platform of Mono- and Polymicrobial Biofilms for High-Throughput Downstream Applications
نویسندگان
چکیده
Current in vitro techniques for the culture of microorganisms, and particularly of delicate microbial biofilms, are still mostly limited to low-density plates and manual labor and are not amenable to automation and true high-throughput (HT) applications. We have developed a novel fully automated platform for the formation of mono- and polymicrobial biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans at the nanoscale level. The nBioChip is robotically printed, robustly handled, and scanned using a standard microarray reader. Using this technique, hundreds to thousands of identical nanobiofilms encapsulated in hydrogel spots were cultured on microscope slides. The spots can withstand the washing steps involved in screening assays. The miniaturized biofilms demonstrated characteristics similar to those displayed by conventionally formed macroscopic biofilms, including (i) three-dimensional architectural features, (ii) synthesis of exopolymeric matrix material, and (iii) elevated resistance to antibiotic treatment. On the basis of our results, the nBioChip can generate reliable high-throughput antimicrobial susceptibility testing (HT-AST) in 12 to 18 h. The chip serves as a proof-of-concept universal platform for high-throughput drug screening and other downstream applications and furthers understanding of microbial interactions in mixed-species communities at the nanoscale level. IMPORTANCE With an estimated 80% of infections being associated with a biofilm mode of growth and the ensuing recalcitrance of these biofilms with respect to conventional antibiotic treatment leading to high mortality rates, there is a dire and unmet need for the development of novel approaches to prevent, treat, and control these infections. Both bacteria and fungi are capable of forming biofilms that are inherently fragile and often polymicrobial in nature, which further complicates treatment. In this work, we showcase a nanobiofilm chip as a convenient platform for culturing several hundreds of mono- or polymicrobial biofilms and for susceptibility testing. This platform enables true ultra-high-throughput screening for antimicrobial drug discovery or diagnostics or for addressing fundamental issues in microbiology.
منابع مشابه
Application Mapping onto Network-on-Chip using Bypass Channel
Increasing the number of cores integrated on a chip and the problems of system on chips caused to emerge networks on chips. NoCs have features such as scalability and high performance. NoCs architecture provides communication infrastructure and in this way, the blocks were produced that their communication with each other made NoC. Due to increasing number of cores, the placement of the cores i...
متن کاملHigh throughput automated chromatin immunoprecipitation as a platform for drug screening and antibody validation.
Chromatin immunoprecipitation (ChIP) is an assay for interrogating protein-DNA interactions that is increasingly being used for drug target discovery and screening applications. Currently the complexity of the protocol and the amount of hands-on time required for this assay limits its use to low throughput applications; furthermore, variability in antibody quality poses an additional obstacle i...
متن کاملA high-throughput microfluidic single-cell screening platform capable of selective cell extraction.
Microfluidic devices and lab-on-a-chip technologies have been extensively used in high-throughput single-cell analysis applications using their capability to precisely manipulate cells as well as their microenvironment. Although significant technological advances have been made in single-cell capture, culture, and analysis techniques, most microfluidic systems cannot selectively retrieve sample...
متن کاملHigh-throughput dental biofilm growth analysis for multiparametric microenvironmental biochemical conditions using microfluidics.
Dental biofilm formation is not only a precursor to tooth decay, but also induces more serious systematic health problems such as cardiovascular disease and diabetes. Understanding the conditions promoting colonization and subsequent biofilm development involving complex bacteria coaggregation is particularly important. In this paper, we report a high-throughput microfluidic 'artificial teeth' ...
متن کاملProteome Profiling by Multidimensional Protein Separation and Automated Lab-on-a-Chip Technology
Technologies and methodologies for proteomic studies often involve a combination of protein fractionation, identification, and quantitation by liquid chromatography tandem mass spectrometry (LC/MS/MS) and in many cases pattern comparison of mass spectra from a complex sample. A common goal of these efforts is the discovery and validation of biomarkers. While current mass spectrometry-based prot...
متن کامل