Shangsheng Feng

Sensitive biomolecule detection in lateral flow assay with a portable temperature–humidity control device

Lateral flow assays (LFAs) have currently attracted broad interest for point-of-care (POC) diagnostics, but their application has been restricted by poor quantification and limited sensitivity. While the former has been currently solved to some extent by the development of handheld or smartphone-based readers, the latter has not been addressed fully, particularly the potential influences of environmental conditions (e.g., temperature and relative humidity (RH)), which have not yet received serious attention. The present study reports the use of a portable temperature-humidity control device to provide an optimum environmental requirement for sensitivity improvement in LFAs, followed by quantification by using a smartphone. We found that a RH beyond 60% with temperatures of 55-60°C and 37-40°C produced optimum nucleic acid hybridization and antigen-antibody interaction in LFAs, respectively representing a 10-fold and 3-fold signal enhancement over ambient conditions (25°C, 60% RH). We envision that in the future the portable device could be coupled with a fully integrated paper-based sample-to-answer biosensor for sensitive detection of various target analytes in POC settings.

Improved sensitivity of lateral flow assay using paper-based sample concentration technique

Lateral flow assays (LFAs) hold great promise for point-of-care testing, especially in resource-poor settings. However, the poor sensitivity of LFAs limits their widespread applications. To address this, we developed a novel device by integrating dialysis-based concentration method into LFAs. The device successfully achieved 10-fold signal enhancement in Human Immunodeficiency Virus (HIV) nucleic acid detection with a detection limit of 0.1 nM and 4-fold signal enhancement in myoglobin (MYO) detection with a detection limit of 1.56 ng/mL in less than 25 min. This simple, low-cost and portable integrated device holds great potential for highly sensitive detection of various target analytes for medical diagnostics, food safety analysis and environmental monitoring.

Polydimethylsiloxane-Paper Hybrid Lateral Flow Assay for Highly Sensitive Point-of-Care Nucleic Acid Testing

In nucleic acid testing (NAT), gold nanoparticle (AuNP)-based lateral flow assays (LFAs) have received significant attention due to their cost-effectiveness, rapidity, and the ability to produce a simple colorimetric readout. However, the poor sensitivity of AuNP-based LFAs limits its widespread applications. Even though various efforts have been made to improve the assay sensitivity, most methods are inappropriate for integration into LFA for sample-to-answer NAT at the point-of-care (POC), usually due to the complicated fabrication processes or incompatible chemicals used. To address this, we propose a novel strategy of integrating a simple fluidic control strategy into LFA. The strategy involves incorporating a piece of paper-based shunt and a polydimethylsiloxane (PDMS) barrier to the strip to achieve optimum fluidic delays for LFA signal enhancement, resulting in 10-fold signal enhancement over unmodified LFA. The phenomena of fluidic delay were also evaluated by mathematical simulation, through which we found the movement of fluid throughout the shunt and the tortuosity effects in the presence of PDMS barrier, which significantly affect the detection sensitivity. To demonstrate the potential of integrating this strategy into a LFA with sample-in-answer-out capability, we further applied this strategy into our prototype sample-to-answer LFA to sensitively detect the Hepatitis B virus (HBV) in clinical blood samples. The proposed strategy offers great potential for highly sensitive detection of various targets for wide application in the near future.

Advances in paper-based sample pretreatment for point-of-care testing

Abstract In recent years, paper-based point-of-care testing (POCT) has been widely used in medical diagnostics, food safety and environmental monitoring. However, a high-cost, time-consuming and equipment-dependent sample pretreatment technique is generally required for raw sample processing, which are impractical for low-resource and disease-endemic areas. Therefore, there is an escalating demand for a cost-effective, simple and portable pretreatment technique, to be coupled with the commonly used paper-based assay (e.g. lateral flow assay) in POCT. In this review, we focus on the importance of using paper as a platform for sample pretreatment. We firstly discuss the beneficial use of paper for sample pretreatment, including sample collection and storage, separation, extraction, and concentration. We highlight the working principle and fabrication of each sample pretreatment device, the existing challenges and the future perspectives for developing paper-based sample pretreatment technique.

High-Throughput Non-Contact Vitrification of Cell-Laden Droplets Based on Cell Printing.

Cryopreservation is the most promising way for long-term storage of biological samples e.g., single cells and cellular structures. Among various cryopreservation methods, vitrification is advantageous by employing high cooling rate to avoid the formation of harmful ice crystals in cells. Most existing vitrification methods adopt direct contact of cells with liquid nitrogen to obtain high cooling rates, which however causes the potential contamination and difficult cell collection. To address these limitations, we developed a non-contact vitrification device based on an ultra-thin freezing film to achieve high cooling/warming rate and avoid direct contact between cells and liquid nitrogen. A high-throughput cell printer was employed to rapidly generate uniform cell-laden microdroplets into the device, where the microdroplets were hung on one side of the film and then vitrified by pouring the liquid nitrogen onto the other side via boiling heat transfer. Through theoretical and experimental studies on vitrification processes, we demonstrated that our device offers a high cooling/warming rate for vitrification of the NIH 3T3 cells and human adipose-derived stem cells (hASCs) with maintained cell viability and differentiation potential. This non-contact vitrification device provides a novel and effective way to cryopreserve cells at high throughput and avoid the contamination and collection problems.

An In Vitro and Numerical Study of Moxibustion Therapy on Biological Tissue

Objective: Moxibustion therapy achieves satisfactory therapeutic effects largely depending on the heat stimulation of burning moxa. Understanding the thermal characteristics of heating process is an effective way to reveal the underlying mechanisms of moxibustion therapy. Methods: This paper performs experimental study on temperature distributions of burning moxa sticks and fresh in vitro porcine abdominal tissue using an infrared camera and thermocouples. Meanwhile, a moxibustion model incorporating moxa stick burning model and tissue heat transfer model was established with consideration of radiation propagation and water evaporation. Results: The burning features of moxa sticks were acquired and the radiation energy generated by the burning moxa stick was absorbed and scattered in biological tissue, resulting in a large temperature gradient in the skin layer. And the water evaporation led to a mass loss and reduced skin surface temperature. The numerical model was verified by experimental results and the effects of moxibustion treatment distance and duration can be quantified based on model calculation. Conclusion: The detailed heat transfer process of moxibustion was obtained experimentally and numerically. During moxibustion, the radiation attenuation and water evaporation have a significant influence on the energy transport in biological tissue which cannot be ignored. The treatment distance of 3 cm is the recommended value to achieve the treatment efficacy without thermal damage and pain. Significance: This research would reveal the underlying mechanisms of moxibustion therapy. Besides, the developed models are expected to establish a guideline for moxibustion clinical treatment.

Thermal Mapping Using Infrared Thermography

Infrared (IR) thermometry is one of the noninvasive temperature measurement techniques allowing surface temperatures both at a point and over an area to be obtained. Such IR thermometry systems (eg, IR camera) are commercially available and integrated with built-in software and a real-time display unit. The following questions arise: (1) Why do readers need to know the basic principles of the IR system? (2) Why is there a need for calibration for a presumably ready-to-go system? The former is to acquaint the readers with how the IR camera works. The latter is dependent upon how close your experimental setup is to the manufacturer’s calibration conditions. This procedure is essential to improve the data accuracy of an IR system in a specific experimental setup whose conditions typically differ from the manufacturers’. A brief introduction to the theoretical background of how the IR system (camera) works is given and some parameters to be considered during calibration are discussed. Examples that describe the necessary details of how to implement the IR thermography are presented.