Colorectal cancer (CRC) is a prevalent and potentially fatal disease categorized based on its high incidences and mortality rates, which raised the need for effective diagnostic strategies for the early detection and management of CRC. While there are several conventional cancer diagnostics available, they have certain limitations that hinder their effectiveness. Significant research efforts are currently being dedicated to elucidating novel methodologies that aim at comprehending the intricate molecular mechanism that underlies CRC. Recently, microfluidic diagnostics have emerged as a pivotal solution, offering non-invasive approaches to real-time monitoring of disease progression and treatment response. Microfluidic devices enable the integration of multiple sample preparation steps into a single platform, which speeds up processing and improves sensitivity. Such advancements in diagnostic technologies hold immense promise for revolutionizing the field of CRC diagnosis and enabling efficient detection and monitoring strategies. This article elucidates several of the latest developments in microfluidic technology for CRC diagnostics. In addition to the advancements in microfluidic technology for CRC diagnostics, the integration of artificial intelligence (AI) holds great promise for further enhancing diagnostic capabilities. Advancements in microfluidic systems and AI-driven approaches can revolutionize colorectal cancer diagnostics, offering accurate, efficient, and personalized strategies to improve patient outcomes and transform cancer management.

This work highlights recent advancements in the integration of analytical tools into DMF devices, and discusses the current challenges and potential solutions as well as future outlooks for an automated, integrative platform for POC applications.

Single-cell sequencing is a powerful technology that provides the capability of analyzing a single cell within a population. This technology is mostly coupled with microfluidic systems for controlled cell manipulation and precise fluid handling to shed light on the genomes of a wide range of cells. So far, single-cell sequencing has been focused mostly on human cells due to the ease of lysing the cells for genome amplification. The major challenges that bacterial species pose to genome amplification from single cells include the rigid bacterial cell walls and the need for an effective lysis protocol compatible with microfluidic platforms. In this work, we present a lysis protocol that can be used to extract genomic DNA from both gram-positive and gram-negative species without interfering with the amplification chemistry. Corynebacterium glutamicum was chosen as a typical gram-positive model and Nostoc sp. as a gram-negative model due to major challenges reported in previous studies. Our protocol is based on thermal and chemical lysis. We consider 80% of single-cell replicates that lead to >5 ng DNA after amplification as successful attempts. The protocol was directly applied to Gloeocapsa sp. and the single cells of the eukaryotic Sphaerocystis sp. and achieved a 100% success rate.

The dynamic immune response to various diseases and therapies has been considered a promising indicator of disease status and therapeutic effectiveness. For instance, the human peripheral blood mononuclear cell (PBMC), as a major player in the immune system, is an important index to indicate a patient’s immune function. Therefore, establishing a simple yet sensitive tool that can frequently assess the immune system during the course of disease and treatment is of great importance. This study introduced an integrated system that includes an electrochemical impedance spectroscope (EIS)-based biosensor in a digital microfluidic (DMF) device, to quantify the PBMC abundance with minimally trained hands. Moreover, we exploited the unique droplet manipulation feature of the DMF platform and conducted a dynamic cell capture assay, which enhanced the detection signal by 2.4-fold. This integrated system was able to detect as few as 10⁴ PBMCs per mL, presenting suitable sensitivity to quantify PBMCs. This integrated system is easy-to-operate and sensitive, and therefore holds great potential as a powerful tool to profile immune-mediated therapeutic responses in a timely manner, which can be further evolved as a point-of-care diagnostic device to conduct near-patient tests from blood samples.

Nucleic acid amplification tests (NAAT) have long been used in laboratory facilities and recently revolutionized the field of molecular diagnostics in point-of-care testing. Digital microfluidics (DMF) has emerged as a promising tool to complete the entire NAAT workflow in a miniaturized format with minimum human intervention. Based on electric fields to manipulate independent reaction droplets, the compact DMF system could perform multiple processes simultaneously and automatically in a programmable fashion. This combination is beginning to establish powerful sample-to-answer platforms in remote or resource-limited settings. Herein, we provide a comprehensive overview of the state-of-the-art DMF technology for point-of-care NAAT. This review focused on key principles of DMF platforms and the latest trends in system integration for automated processes of nucleic acid extraction, amplification, and detection. Also, this article discusses current challenges, including control systems, scalability and throughput, as well as future prospects of DMF-based NAAT strategy for the next generation of point-of-care diagnostics.

Microbial cells behave differently in colonies and when singled out. The standard methods of microbiome studies present the average characteristics and behaviors of heterogeneous populations and lack the resolution to analyze microbes on a single cell level. Besides, the microbiome does not exist in pure colonies in human bodies, but naturally in complex communities. Therefore, it would be ideal to observe the microbes on a single cell level while maintaining their natural settings. Conventional dilution-to-extinction methods are capable of reducing the complexity of the communities to a minimal ecologically functional unit; however, new tools are needed to perform these studies. Recently, microfluidics as a new technology is gaining attention for various single cell applications because it offers the unique ability of handling nanoscale volume in microstructures, providing an attractive alternative to look at single microbial cells. Here, we compare different microfluidic technologies for microbial single cell studies and review the advancement of microfluidics for various microbial single cell analyses. Continuous microfluidics has been used in microbial single cell culture, whole genome sequencing, gene expression, and metabolic analyses; however, droplet microfluidics is becoming a new trend for its high-throughput single cell encapsulation ability. We envision that different microfluidic paradigms will be integrated in the future for their unique attributes and offer a versatile platform for systematic microbiome studies.