Abstract
Fluorescence microscopy can be a powerful tool for cell-based diagnostic assays; however, imaging can be time consuming and labor intensive to perform. Tunable systems give the ability to electronically focus at user selected depths inside an object volume and may simplify the opto-mechanical design of the imaging system. We present a prototype of a universal, tunable, miniature fluorescence microscope built from poly(methyl methacrylate) singlets that incorporates miniature, electrowetted lenses for electronic focusing. We demonstrate the ability of this system to perform clinically relevant differential white blood cell counts using single use custom cartridges pre-loaded with the fluorescent dye acridine orange.
Highlights
Diagnostic tests are essential for rapid and effective health management for a wide range of diseases, from parasitic infections like malaria to infectious diseases like tuberculosis [1,2]
We designed and fabricated a prototype universal, tunable, miniature fluorescence microscope which can operate in the 525-720 nm spectral range
Our system is characterized by a simple opto-mechanical design with all static singlet lenses made exclusively from poly(methyl methacrylate) (PMMA)
Summary
Diagnostic tests are essential for rapid and effective health management for a wide range of diseases, from parasitic infections like malaria to infectious diseases like tuberculosis [1,2]. Fluorescence microscopes can be used to quantify specific reaction products by capturing them on fluorescent beads or to quantify the number of white blood cells (WBC) within a patient sample; our lab in particular works on microscopy based POC systems These systems do not require high resolution or contrast; instead, they rely on target counting (for example, beads or cells) and measuring the color and intensity of fluorescence. Because these systems have low numerical apertures (NA) and low magnifications, they can be designed with large FOV, which allows a single image to include a sufficient number of targets necessary for statistical analysis. We apply the developed prototype to WBC differential counting, as this test may provide rapid, crucial information regarding infection type and condition of a patient
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