Abstract
Instruments that allow the detection of fluorescence signal are invaluable tools for biomedical and clinical researchers. The technique is widely used in cell biology to microscopically detect target proteins of interest in mammalian cells. Importantly, fluorescence microscopy finds major applications in cancer biology where cancer cells are chemically labelled for detection. However, conventional fluorescence detection instruments such as fluorescence imaging microscopes are expensive, not portable and entail potentially high maintenance costs. Here we describe the design, development and applicability of a low-cost and portable fluorometer for the detection of fluorescence signal emitted from a model breast cancer cell line, engineered to stably express the green fluorescent protein (GFP). This device utilizes a flashlight which works in the visible range as an excitation source and a photodiode as the detector. It also utilizes an emission filter to mainly allow the fluorescence signal to reach the detector while eliminating the use of an excitation filter and dichroic mirror, hence, making the device compact, low-cost, portable and lightweight. The custom-built sample chamber is fabricated with a 3D printer to house the detector circuitry. We demonstrate that the developed fluorometer is able to distinguish between the cancer cell expressing GFP and the control cell. The fluorometer we developed exhibits immense potential for future applicability in the selective detection of fluorescently-labelled breast cancer cells.
Highlights
Fluorescence microscopy which is based on the principle of fluorescence imaging is a powerful tool used by many biologists as well as chemists to monitor cell dynamics and molecules in the field of biology and chemistry
In order to evaluate the minimum level of confluency that is required for the device to successfully determine if the cultured cells were control cells or the cells with green fluorescent protein (GFP), the cells of varying confluency were cultured on different coverslips and were seeded on the glass slide
The device was able to detect the cells that were less than 60% confluent, the obtained readings from fluorometer were variable and the difference between the cancer cell with GFP and without GFP was less than 700mV
Summary
Fluorescence microscopy which is based on the principle of fluorescence imaging is a powerful tool used by many biologists as well as chemists to monitor cell dynamics and molecules in the field of biology and chemistry. These commercial fluorescent microscopes offer various advantages: live cell imaging, wide field-of-view, sensitive sophisticated cameras for high resolution images, etc. The fluorescently labelled proteins of interest help researchers to investigate any cellular process and aid in extracting meaningful information These microscopes [1,2] are expensive as they utilize sophisticated optics making it affordable only by hospitals/research lab with healthy financial resources. A few low-cost fluorescent microscopes [4,5,6] were developed to make it accessible to a greater audience from developing countries but low resolution hinders their application
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