Abstract
The use of micrometer-sized droplets for chemical and biochemical analysis has been widely explored. Photolithography is mainly used to fabricate microfluidic devices, which is often employed to form monodisperse microdroplets. Although photolithography enables precise microfabrication, it is not readily available to biochemists because it requires specialized equipment such as clean room and mask aligners, and expensive consumables such as photoresist and silicon wafers. In this study, we fabricated a microfluidic device using a consumer laser cutter and applied it to droplet formation. Monodisperse microdroplets were formed by using an oil phase for droplet digital polymerase chain reaction (PCR) as the continuous phase and phosphate-buffered saline or polyethylene glycol solution as the dispersed phase. The droplet size decreased as the flow rate of the continuous phase increased and approached a constant value. The method developed in this study can be used to realize microdroplet-based biochemical analysis with simple devices or to construct artificial cells.
Highlights
Many chemical and biochemical analyses using microdroplets have been reported [1,2,3,4]
The typical application of microdroplets is droplet digital polymerase chain reaction [5], in which a sample solution of unknown concentration is diluted with reagents and divided into a thousand of microdroplets, and each droplet contains one molecule of the target DNA or none at all
When polymerase chain reaction (PCR) is performed in this situation, DNA is amplified only in the droplet containing the target DNA, and fluorescence is observed when the fluorescent dye binds to the amplified DNA
Summary
Many chemical and biochemical analyses using microdroplets have been reported [1,2,3,4]. This is a method toisform by dividing the dispersed phase the continuous phase [15,16 tolithography) one droplets of the main fabrication methods forbymicrofluidic devices using micrometer-scale channels This technique enables accurate microfabrication, it requires expensiv the solutions at a constant flow rate using a syringe pump to maintain the shear force acting and extensive facilities and equipment such as clean rooms and mask aligners, expensiv on the dispersed phase at a constant level. Current laser-based various microfabrication methods have been reported, but, as reviewedmicrofabricatio recently methods remain complicated and often require expensive machines [22] Considering th in this journal by Puryear III [22], it is important for beginners to utilize the fabrication aforementioned characteristics of laser-based methods, methods it is clear that bi process with high familiarity. Figure illustration of microdroplet formation in a T-shaped microfluidic channel. channel
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