Music Box-Inspired Semi-Automatic Hematocrit Validation Device.

A high hematocrit (HCT) level is strongly associated with the risk of cardiovascular disease. For early diagnosis of cardiovascular disease, it is vital to regularly measure the HCT, which is typically achieved by centrifuging a blood sample to measure the percentage of red blood cells. However, the centrifugal modalities are usually bulky, expensive, and require a stable electric input, which restrict the availability. This research develops a semi-automatic and portable centrifugal device for HCT measurement. This torque-actuated semi-automatic centrifuge, which we call the tFuge, is inspired by a music box, allowing different operators to generate the same rhythm. It is electricity-free and can be controlled based on a constant torque mechanism. Repeatable test results can be received from among different users regardless of their age, sex, and activity. With the assistance of the Boycott effect on the tFuge, we proved that the HCT level is in high linearity to the length of the sedimentation of the blood cells in a tube (R2 = 0.99, sample HCT range 10-60%). The tFuge takes less than 4 min and requires no more than 10 µL of blood that can be obtained by a less-invasive finger prick to complete the testing procedure. Calibrated gradient numbers are printed onto the rotation disc for instant HCT results that can be read by the naked eye. We expect this proposed point-of-care testing device possesses the potential to replace the microhematocrit centrifuge in the regions with limited resources.

Hand-Powered Microfluidics for Parallel Droplet Digital Loop-Mediated Isothermal Amplification Assays.

Droplet digital loop-mediated isothermal amplification (ddLAMP) is an important assay for pathogen detection due to its high accuracy, specificity, and ability to quantify nucleic acids. However, performing ddLAMP requires expensive instrumentation and the need for highly trained personnel with expertise in microfluidics. To make ddLAMP more accessible, a ddLAMP assay is developed, featuring significantly decreased operational difficulty and instrumentation requirements. The proposed assay consists of three simplified steps: (1) droplet generation step, in which a LAMP mixture can be emulsified just by manually pulling a syringe connected to a microfluidic device. In this step, for the first time, we verify that highly monodispersed droplets can be generated with unstable flow rates or pressures, allowing untrained personnel to operate the microfluidic device and perform ddLAMP assay; (2) heating step, in which the droplets are isothermally heated in a water bath, which can be found in most laboratories; and (3) result analysis step, in which the ddLAMP result can be determined using only a fluorescence microscopy and an open-source analyzing software. Throughout the process, no droplet microfluidic expertise or equipment is required. More importantly, the proposed system enables multiple samples to be processed simultaneously with a detection limit of 10 copies/µL. The test is simple and intuitive to operate in most laboratories for multi-sample detection, significantly enhancing the accessibility and detection throughput of the ddLAMP technique.

An electricity- and instrument-free infectious disease sensor based on a 3D origami paper-based analytical device.

Infectious diseases cause millions of deaths annually in the developing world. Recently, microfluidic paper-based analytical devices (µPADs) have been developed to diagnose such diseases, as these tests are low cost, biocompatible, and simple to fabricate. However, current µPADs are difficult to use in resource-limited areas due to their reliance on external instrumentation to measure and analyze the test results. In this work, we propose an electricity and external instrumentation-free µPAD sensor based on the colorimetric enzyme-linked immunosorbent assay (ELISA) for the diagnosis of infectious disease (3D-tPADs). Designed based on the principle of origami, the proposed µPAD enables the sequential steps of the colorimetric ELISA test to be completed in just ∼10 min. In addition, in order to obtain an accurate ELISA result without using any instrument, we have integrated an electricity-free "timer" within the µPAD that can be controlled by the buffer viscosity and fluid path volume to indicate the appropriate times for washing and color development steps, which can avoid false positive or false negative results caused by an extended or shortened amount of washing and development times. Due to the low background noise and high positive signal intensity of the µPAD, positive and negative detection results can be distinguished by just the naked eye. Furthermore, the ELISA result can be semi-quantified by comparing the results shown on the µPAD with a color chart diagram with a detection limit of HIV type 1(HIV-1) p24 antigen as low as 0.03 ng mL-1. These results demonstrate the proposed sensor can perform infectious disease diagnosis without external instrumentation or electricity, extending the application of the µPAD test for on-site detection and use in resource-limited settings.