Ventilation-Based Strategy to Manage Intraoperative Aerosol Viral Transmission in the Era of SARS-CoV-2.

In operating theaters, ventilation systems are designed to protect the patient from airborne contamination for minimizing risks of surgical site infections (SSIs). Ventilation systems often produce an airflow pattern that continuously pushes air out of the area surrounding the operating table, and hence reduces the resident time of airborne pathogen-carrying particles at the patient's location. As a result, patient-released airborne particles due to the use of powered tools, such as surgical smoke and insufflated CO2, typically circulate within the room. This circulation exposes the surgical team to airborne infection-especially when operating on a patient with infectious diseases, including COVID-19. This study examined the flow pattern of functional ventilation configurations in view of developing ventilation-based strategies to protect both the patient and the surgical team from aerosolized infections. A favorable design that minimized particle circulation was deduced using experimentally validated numerical models. The parameters adapted to quantify circulation of airborne particles were particles' half-life and elevation. The results show that the footprint of the outlet ducts and resulting flow pattern are important parameters for minimizing particle circulation. Overall, this study presents a modular framework for optimizing the ventilation systems that permits a switch in operation configuration to suit different operating procedures.

Affinity-Based Microfluidics Combined with Atomic Force Microscopy for Isolation and Nanomechanical Characterization of Circulating Tumor Cells.

In this chapter, we present the materials and methods required to isolate and characterize circulating tumor cells (CTCs) from blood samples of cancer patients based on our newly developed microfluidic technologies. In particular, the devices presented herein are designed to be compatible with at\omic force microscopy (AFM) for post-capture nanomechanical investigation of CTCs. Microfluidics is well-established as a technology for isolating CTCs from the whole blood of cancer patients, and AFM is a gold standard for quantitative biophysical analysis of cells. However, CTCs are very scarce in nature, and those captured using standard closed-channel microfluidic chips are typically inaccessible for AFM procedures. As a result, their nanomechanical properties largely remain unexplored. Thus, given limitations associated with current microfluidic designs, significant efforts are put toward bringing innovative designs for real time characterization of CTCs. In light of this constant endeavor, the scope of this chapter is to compile our recent efforts on two microfluidic technologies, namely, the AFM-Chip and the HB-MFP, which proved to be efficient in isolating CTCs through antibody-antigen interactions, and their subsequent characterization using AFM.

Characterizing circulating tumor cells using affinity-based microfluidic capture and AFM-based biomechanics.

Elasticity and bio-adhesiveness of circulating tumor cells (CTCs) are important biomarkers of cancer. CTCs are rare in blood, thus their capture and atomic force microscopy (AFM)-based biomechanical characterization require use of multifunctional microfluidic device. Here, we describe procedures for fabrication of such device, AFM-Chip, and give details on its use in affinity-based CTC capture, and integration with AFM via reversable physical assembly. In the AFM-Chip, CTC capture is efficient, and transition to AFM characterization is seamless with minimal cell loss. For complete details on the use and execution of this protocol, please refer to Deliorman et al. (2020).

3D Generation of Multipurpose Atomic Force Microscopy Tips.

In this work, 3D polymeric atomic force microscopy (AFM) tips, referred to as 3DTIPs, are manufactured with great flexibility in design and function using two-photon polymerization. With the technology holding a great potential in developing next-generation AFM tips, 3DTIPs prove effective in obtaining high-resolution and high-speed AFM images in air and liquid environments, using common AFM modes. In particular, it is shown that the 3DTIPs provide high-resolution imaging due to their extremely low Hamaker constant, high speed scanning rates due to their low quality factor, and high durability due to their soft nature and minimal isotropic tip wear; the three important features for advancing AFM studies. It is also shown that refining the tip end of the 3DTIPs by focused ion beam etching and by carbon nanotube inclusion substantially extends their functionality in high-resolution AFM imaging, reaching angstrom scales. Altogether, the multifunctional capabilities of 3DTIPs can bring next-generation AFM tips to routine and advanced AFM applications, and expand the fields of high speed AFM imaging and biological force measurements.

AFM-compatible microfluidic platform for affinity-based capture and nanomechanical characterization of circulating tumor cells.

Circulating tumor cells (CTCs) carried by the patient's bloodstream are known to lead to the metastatic spread of cancer. It is becoming increasingly clear that an understanding of the nanomechanical characteristics of CTCs, such as elasticity and adhesiveness, represents advancements in tracking and monitoring cancer progression and metastasis. In the present work, we describe a combined microfluidic-atomic force microscopy (AFM) platform that uses antibody-antigen capture to routinely isolate and nanomechanically characterize CTCs present in blood samples from prostate cancer patients. We introduce the reversible assembly of a microfluidic device and apply refined and robust chemistry to covalently bond antibodies onto its glass substrate with high density and the desired orientation. As a result, we show that the device can efficiently capture CTCs from patients with localized and metastatic prostate cancer through anti-EpCAM, anti-PSA, and anti-PSMA antibodies, and it is suitable for AFM measurements of captured intact CTCs. When nanomechanically characterized, CTCs originating from metastatic cancer demonstrate decreased elasticity and increased deformability compared to those originating from localized cancer. While the average adhesion of CTCs to the AFM tip surface remained the same in both the groups, there were fewer multiple adhesion events in metastatic CTCs than there were in their counterparts. The developed platform is simple, robust, and reliable and can be useful in the diagnosis and prognosis of prostate cancer as well as other forms of cancer.

An integrated adipose-tissue-on-chip nanoplasmonic biosensing platform for investigating obesity-associated inflammation.

Although many advanced biosensing techniques have been proposed for cytokine profiling, there are no clinically available methods that integrate high-resolution immune cell monitoring and in situ multiplexed cytokine detection together in a biomimetic tissue microenvironment. The primary challenge arises due to the lack of suitable label-free sensing techniques and difficulty for sensor integration. In this work, we demonstrated a novel integration of a localized-surface plasmon resonance (LSPR)-based biosensor with a biomimetic microfluidic 'adipose-tissue-on-chip' platform for an in situ label-free, high-throughput and multiplexed cytokine secretion analysis of obese adipose tissue. Using our established adipose-tissue-on-chip platform, we were able to monitor the adipose tissue initiation, differentiation, and maturation and simulate the hallmark formation of crown-like structures (CLSs) during pro-inflammatory stimulation. With integrated antibody-conjugated LSPR barcode sensor arrays, our platform enables simultaneous multiplexed measurements of pro-inflammatory (IL-6 and TNF-α) and anti-inflammatory (IL-10 and IL-4) cytokines secreted by the adipocytes and macrophages. As a result, our adipose-tissue-on-chip platform is capable of identifying stage-specific cytokine secretion profiles from a complex milieu during obesity progression, highlighting its potential as a high-throughput preclinical readout for personalized obesity treatment strategies.