A microfluidic device is described, demonstrating its fabrication and operation for the capture of single DNA molecules within chambers, enabling tumor-specific biomarker detection via a passive geometric strategy.

For biological and medical research, the non-invasive collection of target cells, like circulating tumor cells (CTCs), is essential. Complex procedures are frequently employed for conventional cell collection, entailing either size-differentiated sorting or invasive enzymatic reactions. We present a functional polymer film, which incorporates the thermoresponsive polymer poly(N-isopropylacrylamide) and the conducting polymer poly(34-ethylenedioxythiopene)/poly(styrene sulfonate), and its utility in the capture and release processes of circulating tumor cells. The proposed polymer films, when applied to microfabricated gold electrodes, are capable of noninvasively capturing and controlling the release of cells, enabling monitoring of these processes concurrently with conventional electrical measurements.

For the creation of new, innovative microfluidic in vitro platforms, stereolithography-based additive manufacturing (3D printing) provides a beneficial approach. This manufacturing method expedites production time, allows for quick alterations to designs, and makes the construction of complex, unified structures possible. The platform, outlined in this chapter, is designed for the capture and evaluation of cancer spheroids maintained in perfusion. Within a workflow involving 3D Petri dish culture, staining, loading, and subsequent imaging under dynamic flow conditions, spheroids are incorporated into 3D-printed devices. This design's implementation of active perfusion enables prolonged viability within intricate 3D cellular constructs, producing results that mirror in vivo conditions far better than those obtained from static monolayer cultures.

The involvement of immune cells in cancer is multifaceted, encompassing their ability to restrain tumor formation by releasing pro-inflammatory signaling molecules, as well as their role in promoting tumor development through the secretion of growth factors, immunosuppressants, and enzymes that modify the extracellular environment. In conclusion, the ex vivo examination of the secretory function of immune cells establishes it as a credible prognostic indicator in cancer. Yet, a critical impediment in present methods to investigate the ex vivo secretion function of cells is their low processing rate and the significant consumption of sample material. Microfluidics's integration capability of components, including cell culture and biosensors, within a monolithic microdevice is a unique strength; this capability maximizes analytical throughput and leverages the inherent reduced sample requirements. Moreover, automated analysis of this kind is facilitated by the integration of fluid control elements, thereby improving the consistency of results. Employing a highly integrated microfluidic device, we describe an approach to analyze the ex vivo secretory function of immune cells.

Circulating tumor cell (CTC) clusters, exceptionally rare and found in the bloodstream, can be isolated for minimally invasive diagnostic and prognostic purposes, revealing their contribution to metastasis. Specific technologies designed to improve CTC cluster enrichment prove inadequate in terms of practical processing speed for clinical implementation, or their design can cause potentially harmful high shear forces, leading to the disintegration of large clusters. Biochemistry and Proteomic Services We have developed a methodology for the rapid and effective isolation of CTC clusters from cancer patients, irrespective of cluster size or cell surface marker profile. Cancer screening and personalized medicine will fundamentally incorporate the minimally invasive access to tumor cells found within the hematogenous circulation.

The nanoscopic bioparticles, small extracellular vesicles (sEVs), facilitate the transport of biomolecular cargo across cellular boundaries. Among numerous pathological processes, electric vehicles have been implicated in some cases, notably cancer, making them promising prospects for development of diagnostic and therapeutic interventions. Characterizing the distinctive protein and RNA content of secreted extracellular vesicles could reveal their influence on cancer progression. Nonetheless, the undertaking faces a challenge stemming from the comparable physical characteristics of sEVs and the necessity for highly discerning analytical procedures. The sEV subpopulation characterization platform (ESCP), a platform using surface-enhanced Raman scattering (SERS) readouts for a microfluidic immunoassay, is detailed in our method of preparation and operation. To enhance the collisions of sEVs with the antibody-functionalized sensor surface, ESCP employs an electrohydrodynamic flow induced by an alternating current. Polyinosinic-polycytidylic acid sodium TLR activator sEVs, captured and labeled with plasmonic nanoparticles, are characterized phenotypically in a multiplexed and highly sensitive fashion using SERS. sEVs (exosomes) derived from cancer cell lines and plasma samples are evaluated for the expression of three tetraspanins (CD9, CD63, CD81) and four cancer-associated biomarkers (MCSP, MCAM, ErbB3, LNGFR) using the ESCP technique.

To determine the grouping of malignant cells detected in blood and other bodily fluids, liquid biopsies are utilized as examination processes. Patient discomfort is considerably minimized with liquid biopsies, a significantly less invasive procedure than tissue biopsies, requiring only a small quantity of blood or body fluids. Fluid biopsies, processed with microfluidic systems, can yield isolated cancer cells for timely diagnosis. Microfluidic devices are finding an expanding application in the ever-evolving field of 3D printing. Compared to traditional microfluidic device production, 3D printing offers several advantages, such as the straightforward creation of numerous precise replicas on a large scale, the incorporation of novel materials, and the execution of intricate or time-consuming procedures difficult to implement with conventional microfluidic devices. genetic clinic efficiency Utilizing 3D printing in conjunction with microfluidics enables a relatively economical approach to liquid biopsy analysis, with the resulting chip surpassing traditional microfluidic designs in usability. A 3D microfluidic chip approach to affinity-based separation of cancer cells from liquid biopsies, and its supporting rationale, are the subject of this chapter's examination.

The increasing focus within oncology is on individualized strategies for determining the effectiveness of a chosen therapy in each patient. The remarkable precision of personalized oncology has the potential to lead to a substantial extension of patient survival times. As a primary source of patient tumor tissue, patient-derived organoids are crucial for therapy testing in personalized oncology. Cancer organoid cultures adhere to the gold standard methodology of utilizing Matrigel-coated multi-well plates. These standard organoid cultures, whilst showing effectiveness, nonetheless present limitations, specifically the requirement for a sizable initial cell population and the varied sizes of the resulting cancer organoids. This secondary hindrance presents obstacles in tracking and assessing variations in organoid dimensions as a consequence of therapy. To both decrease the starting cellular material for organoid formation and standardize organoid sizes for easier therapy assessments, microfluidic devices with integrated microwell arrays can be employed. This paper details the methods for constructing microfluidic devices, cultivating patient-derived cancer cells, developing organoids, and evaluating treatments within these systems.

Bloodstream-circulating tumor cells (CTCs), though few in number, act as a valuable predictor of cancer progression. Obtaining highly purified, intact circulating tumor cells (CTCs) with the desired level of viability is difficult, because they represent a tiny fraction of the blood cell population. This chapter details the construction and implementation of a novel, self-amplified inertial-focused (SAIF) microfluidic chip. This chip facilitates the high-throughput, label-free separation of circulating tumor cells (CTCs) from patient blood, based on their size. The SAIF chip, featured in this chapter, demonstrates the capability of a narrow, zigzag channel (40 meters wide) connected with expansion zones to efficiently sort cells of diverse dimensions, effectively lengthening the separation distance.

Determining the malignancy relies on the identification of malignant tumor cells (MTCs) present in pleural effusions. While the sensitivity of MTC detection is maintained, it is markedly hampered by the substantial number of background blood cells in large-scale samples. An inertial microfluidic sorter coupled with an inertial microfluidic concentrator is presented herein for the on-chip isolation and enrichment of malignant pleural tumor cells (MTCs) from malignant pleural effusions (MPEs). The designed sorter and concentrator exploit intrinsic hydrodynamic forces to position cells at their respective equilibrium points. This capability enables the sorting of cells by size and allows for the removal of cell-free fluids for cell enrichment. This technique permits the near-total elimination of background cells and an exceptionally high, 1400-fold, enrichment of MTCs from large MPE samples. For accurate MPE identification in cytological examinations, immunofluorescence staining can be directly applied to the concentrated and highly pure MTC solution. The suggested method can facilitate both the identification and the enumeration of rare cells across different clinical samples.

Involved in the intricate dance of cell-cell communication are extracellular vesicles, specifically exosomes. Recognizing their bioavailability and presence in all body fluids, including blood, semen, breast milk, saliva, and urine, their use as an alternative, non-invasive method for diagnosing, monitoring, and predicting numerous diseases, such as cancer, has been recommended. The isolation and subsequent analysis of exosomes show promise in the fields of diagnostics and personalized medicine. Differential ultracentrifugation, while a prevalent isolation technique, suffers from significant drawbacks, including labor intensity, extended duration, high costs, and limited yield. Exosome isolation is now facilitated by emerging microfluidic devices, providing a low-cost, high-purity, and rapid method of treatment.