Our approach leverages a microfluidic device employing antibody-functionalized magnetic nanoparticles to capture and separate components from the inflowing whole blood. This device isolates pancreatic cancer-derived exosomes directly from whole blood, thereby achieving high sensitivity, without any pretreatment steps.

Clinical medicine utilizes cell-free DNA, significantly for cancer detection and the oversight of cancer treatment protocols. Microfluidic-based systems promise rapid and economical, decentralized detection of circulating tumor DNA in blood samples, also known as liquid biopsies, eliminating the need for invasive procedures or expensive imaging techniques. This method showcases a straightforward microfluidic system for the extraction of cell-free DNA from 500 microliters of plasma. Either static or continuous flow systems are suitable for this technique, which can also be implemented as a standalone module or incorporated into a lab-on-chip platform. The system's operation depends on a simple yet highly versatile bubble-based micromixer module, with its specialized components potentially created through low-cost rapid prototyping techniques or via readily available 3D-printing services. This system boasts a tenfold improvement in cell-free DNA extraction from small blood plasma samples, surpassing control methods in capture efficiency.

Fine-needle aspiration (FNA) sample diagnostic accuracy from cysts, fluid-filled, potentially precancerous sacs, is significantly boosted by rapid on-site evaluation (ROSE), though this method's effectiveness hinges on cytopathologist expertise and accessibility. A semiautomated sample preparation device for ROSE is demonstrated. A single device incorporates a smearing tool and a capillary-driven chamber to complete the smearing and staining procedures for an FNA sample. A demonstration of the device's ability to prepare samples for ROSE analysis is presented, utilizing a human pancreatic cancer cell line (PANC-1) and FNA samples from the liver, lymph node, and thyroid. This device, engineered using microfluidic principles, decreases the quantity of equipment required for FNA sample preparation within surgical settings, potentially broadening the implementation of ROSE procedures in healthcare institutions.

The analysis of circulating tumor cells, using newly developed enabling technologies, has provided new insights into cancer management in recent years. In spite of their development, most of the implemented technologies are challenged by excessive costs, time-consuming workflows, and a reliance on specialized equipment and operators. biocidal activity A microfluidic device-based workflow for isolating and characterizing single circulating tumor cells is proposed herein. By handling the entire process, a laboratory technician can complete it in just a few hours after sample collection, without any reliance on microfluidic expertise.

The use of microfluidic technologies allows for the production of substantial datasets, while consuming less cellular and reagent material than traditional well plate methodologies. With miniaturized methods, the development of intricate 3-dimensional preclinical models of solid tumors, possessing precisely controlled sizes and cell constitutions, becomes possible. To minimize experimental costs during the development of immunotherapies and combination therapies, recreating the tumor microenvironment for preclinical screening at a scalable level is essential. This process uses physiologically relevant 3D tumor models to evaluate the effectiveness of the therapies. This paper details the manufacturing of microfluidic devices and the subsequent protocols used for cultivating tumor-stromal spheroids, enabling the assessment of anti-cancer immunotherapies' efficacy as single agents or as part of a combined treatment approach.

Using genetically encoded calcium indicators (GECIs) and high-resolution confocal microscopy, the dynamic visualization of calcium signals within cells and tissues is achievable. in vivo pathology The mechanical micro-environments of tumor and healthy tissues are mimicked by programmable 2D and 3D biocompatible materials. Physiologically relevant functions of calcium dynamics within tumors at different stages of progression are revealed through the use of cancer xenograft models and ex vivo functional imaging of tumor slices. The integration of these formidable methods empowers us to quantify, diagnose, model, and understand the intricate pathobiology of cancer. IMP1088 Detailed materials and methods for establishing this integrated interrogation platform are presented, ranging from the generation of transduced cancer cell lines, stably expressing CaViar (GCaMP5G + QuasAr2), to in vitro and ex vivo calcium imaging in 2D/3D hydrogels and tumor tissues. These instruments enable in-depth studies of mechano-electro-chemical network dynamics in biological systems.

Machine learning-powered impedimetric electronic tongues, incorporating nonselective sensors, are expected to bring disease screening biosensors into mainstream clinical practice. These point-of-care diagnostics are designed for swift, precise, and straightforward analysis, potentially rationalizing and decentralizing laboratory testing with considerable social and economic implications. This chapter details the concurrent determination of two extracellular vesicle (EV) biomarkers, namely the concentrations of EVs and their associated protein cargo, in mice blood afflicted with Ehrlich tumors. This is achieved through the combination of a cost-effective and scalable electronic tongue with machine learning, extracting data from a single impedance spectrum without employing biorecognition elements. Manifestations of mammary tumor cells are prominently displayed in this tumor specimen. Microfluidic chips fabricated from polydimethylsiloxane (PDMS) now incorporate HB pencil core electrodes. The platform achieves superior throughput compared to the literature's techniques for quantifying EV biomarkers.

Investigating the molecular hallmarks of metastasis and developing personalized therapies benefits from the selective capture and release of viable circulating tumor cells (CTCs) obtained from the peripheral blood of cancer patients. CTC-based liquid biopsies are gaining significant traction in the clinical sphere, offering clinicians the ability to track patients' real-time responses during clinical trials and improve accessibility to diagnosing cancers that were previously difficult to identify. However, circulating tumor cells (CTCs) are less common than the broader population of cells residing in the circulatory system, leading to the development of new microfluidic device designs. Current microfluidic approaches for circulating tumor cells (CTCs) isolation are frequently plagued by a fundamental dilemma: attaining a substantial increase in circulating tumor cell concentration often comes at a considerable expense to cellular viability, or if viability is maintained, the enrichment of circulating tumor cells is suboptimal. To achieve high-efficiency capture of circulating tumor cells (CTCs) and high cell viability, we introduce a procedure for fabricating and operating a microfluidic device. Utilizing nanointerface-functionalized microvortex-inducing microfluidic devices, circulating tumor cells (CTCs) are effectively enriched via cancer-specific immunoaffinity. Subsequently, a thermally responsive surface chemistry releases the captured cells upon heating to 37 degrees Celsius.

In this chapter, we describe the required materials and methods for the isolation and characterization of circulating tumor cells (CTCs) from cancer patient blood, achieved through our advanced microfluidic technology. Specifically, the devices described here are intended for compatibility with atomic force microscopy (AFM), enabling post-capture nanomechanical investigation of circulating tumor cells (CTCs). Whole blood from cancer patients can be effectively processed via microfluidic methods to isolate circulating tumor cells (CTCs), with atomic force microscopy (AFM) acting as the definitive approach for quantifying the biophysical characteristics of cells. Although circulating tumor cells are present in low numbers in nature, they are often difficult to access for atomic force microscopy (AFM) analysis following capture with standard closed-channel microfluidic systems. Thus, a substantial amount of work remains to be done in understanding their nanomechanical properties. In light of the limitations present in current microfluidic designs, a considerable focus is placed on the development of innovative layouts to facilitate real-time characterization of circulating tumor cells. Considering this ongoing work, this chapter brings together our latest work on two microfluidic platforms, the AFM-Chip and HB-MFP, proving successful in isolating CTCs using antibody-antigen interactions, followed by AFM analysis.

Within the context of precision medicine, the speed and accuracy of cancer drug screening are of significant importance. Nevertheless, the small amount of tumor biopsy specimens has prevented the use of conventional drug screening protocols with microwell plates for each unique patient. A microfluidic device serves as an excellent platform for the meticulous handling of small sample volumes. The evolving platform effectively supports assays concerning nucleic acids and cells. Despite this, the straightforward provision of drugs for on-chip cancer drug screening in clinical trials remains a difficult task. The process of combining droplets with consistent dimensions, adding drugs to attain a desired screened concentration, proved to be significantly more intricate than previous on-chip drug dispensing protocols. A novel digital microfluidic system is introduced here, employing a specially structured electrode (a drug dispenser). High-voltage-driven droplet electro-ejection dispenses drugs with convenient adjustment through external electrical controls. The screened drug concentrations using this system can cover a range up to four orders of magnitude, while maintaining a low sample consumption. The cell sample can receive customized drug dosages via a versatile electric delivery system. Moreover, an on-chip platform allows for quick and easy screening, encompassing either a single drug or a combination of medications.