Abstract
Developing more efficient methods for antibiotic susceptibility testing is a pressing issue in novel drug development as bacterial resistance to antibiotics becomes increasingly common. Microfluidic devices have been demonstrated to be powerful platforms that allow researchers to perform multiplexed antibiotic testing. However, the level of multiplexing within microdevices is limited, evidencing the need of creating simple, low-cost and high-resolution imaging systems that can be integrated in antibiotic development pipelines. This paper describes the design and development of an epifluorescence inverted microscope that enables long-term monitoring of bacteria inside multiplexed microfluidic devices. The goal of this work is to provide a simple microscope powerful enough to allow single-cell analysis of bacteria at a reduced cost. This facilitates increasing the number of microscopes that are simultaneously used for antibiotic testing. We prove that the designed system is able to accurately detect fluorescent beads of 100 nm, demonstrating comparable features to high-end commercial microscopes and effectively achieving the resolution required for single-cell analysis of bacteria. The proposed microscope could thus increase the efficiency in antibiotic testing while reducing cost, size, weight, and power requirements, contributing to the successful development of new antibiotic drugs.
Highlights
Multidrug resistant bacterial diseases have become a major concern worldwide
We have developed an epifluorescence inverted microscope for imaging bacteria inside microfluidic devices during antibiotic susceptibility testing
For antibiotic drug development, multiplexing is pursued to increase the efficiency of the technique, but the level of multiplexing within the microdevice is limited by the area of acquisition of the microscope
Summary
Multidrug resistant bacterial diseases have become a major concern worldwide. Its causes are primarily found in treatment mismanagement, direct transmission of the resistant pathogens, and rapid spontaneous mutation [1]. One of the main problems associated with multidrug bacterial resistance is the fact that, currently, there is not a fast enough diagnostic procedure to guide the prescription of the optimal antibiotic for a specific disease. The standard procedures carried out require taking the patient’s samples to a microbiology lab and waiting several days to obtain a pure culture of the pathogen [2]. A faster approach could involve the use of lab-on-a-chip technology, which could guide antibiotic prescriptions in a much efficient way [3]. Besides solutions at the patient level, new drugs or regimes combining several drugs are essential to tackle new strains of drug-resistant bacteria; Sensors 2020, 20, 4140; doi:10.3390/s20154140 www.mdpi.com/journal/sensors
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