Rapid Prototyping for Detection and Sampling with Applications in Chemistry and Biology Laboratories

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Thursday, March 31, 2022 1:00 pm EST
Organizers: Phillip Mach, US Army + Trevor Glaros, Los Alamos National Laboratory

Utilizing three dimensional printing for rapid prototyping has dramatically redefined what types of experiments are possible in laboratories throughout the world. It is now possible to go from an untested idea to a working prototype without have to navigate or undergo the expense of complex manufacturing processes. Due to this technology’s wide availability, speed, diversity of ‘printable’ materials, and relatively low cost, 3D printing and lithography techniques to be a valuable tool that has facilitated rapid advances in chemistry and biology laboratories worldwide. This symposium aims to highlight some of the most creative advances that sit at this unique interface of material and physical sciences. Applications presented within this symposium range from advances in organ-on-a-chip cell culturing systems to unique approaches for capturing micro aerosols known to spread airborne respiratory viruses. Our hope is the inspire thought provoking ideas in the biological and chemical sciences that may be possible due to the rapid innovations in the additive manufacturing processes.

Presentation 1
Lessons-Learned in Using Rapid Prototyping for Aerosol Sampling and Analysis
Igor Novosselov, University of Washington
Covid-19 pandemic and increased frequency of wildfires have elevated the word “aerosol” to a household term status; however, the aerosol science community had been researching the effects of these airborne particulate matter (PM) on human health and environment. For example, PM emitted from combustion sources impacts the radiative forcing of climate and adversely affects human health, such as pulmonary, cardiovascular, neurological diseases, and cancer. Biological aerosols such as virus and bacterial species are responsible for the spread of airborne diseases, while other biological aerosols can trigger acute toxic responses, exacerbate asthma and allergies in sensitive populations. For decades, aerosol sampling equipment, instrumentation, and analytical methods primarily targeted the research community and environmental health specialists; thus, the form-factor and usability of these equipment and analytical techniques were not considered. With advances in electronics and fabrication methods, a new trend in low-cost, user-friendly devices appeared. This development is reflected in thousands of peer-reviewed publications related to miniature sensors, microfluidics, nanoscale devices. However, to autonomously operate the low-cost devices, the sample still must be delivered to the sensor. This talk will address the benefits and challenges of using rapid prototyping for aerosol sampling and analysis. Two case studies will be described: (1) distributed PM sensor network for monitoring PM in the medical settings and their collection for subsequent identification; (2) capture and in-situ fluorescent spectroscopy analysis of combustion generated aerosols. Fabrication techniques, material selection significantly affect usability, robustness, and data quality. In most cases, rapid prototyping was helpful in the iterative development cycle; however, achieving a reliable long-term performance of aerosol sampling and analysis devices was found to be challenging.

Presentation 2
Quantitative In Vitro Proteomic Profiling of Colon Cancer Spheroids Treated With Combination Chemotherapy
Amanda Hummon, University of Notre Dame
For patients diagnosed with metastatic colorectal cancer, there are limited clinical options. New approaches are needed to identify putative therapies earlier in the drug development process. Here, an in vitro platform was used to assess the treatment of three-dimensional colon cancer spheroids with the combination chemotherapy FOLFIRI (folinic acid, 5-fluorouracil, and irinotecan). The drugs were administered in a dynamic fashion with the use of a custom 3D printed fluidic device. After dosing, the spheroids were harvested for quantitative proteomic profiling by liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) and also assessed with MALDI-Imaging Mass Spectrometry (IMS) to map the spatial distribution of the chemotherapeutics and metabolites. After treatment, the penetration of folinic acid was detected in the core of the spheroids while the metabolites were detected primarily in the outer proliferating rim. Quantitative labeling of proteins revealed the alteration of several cancer-associated pathways. The combination of this dosing regime, combined with quantitative proteomic analysis in a three dimensional cell culture system, is a high-throughput and cost-effective approach for preclinical evaluation of drug candidates prior to animal testing.

Presentation 3
Paper Spray Mass Spectrometry using 3D Printed Devices for Forensic, Clinical, and Environmental Applications
Sarah Dowling, Indiana University-Purdue University Indianapolis
3D printing has become a valuable tool in analytical chemistry. Not only is it a cost-effective manufacturing technique, but devices can be developed to simplify complex analyses that would normally require a trained analyst and lengthy sample preparation. Our work focuses on the exploitation of various 3D printed assemblies to facilitate paper spray mass spectrometry (PS-MS). In PS-MS, a crude biofluid sample is applied to a piece of chromatography paper that is cut to a point. A solvent is used to extract the analytes via capillary action. Then a voltage is applied which facilitates the ionization of the analytes of interest. Three projects will be discussed that utilize 3D printing in different ways. The first project will discuss a 3D printed paper spray cartridge containing an integrated solid phase extraction (SPE) column for the analysis of drugs of abuse in ER patient samples. Low and sub part-per-billion limits of identification in plasma were achieved using this cartridge. The overall assay requires a fraction of the effort compared to traditional SPE extraction and HPLC-MS analysis due to the elimination of extra sample preparation steps. In the second project, a 3D printed cartridge was developed for targeted protein detection. The all-in-one cartridge included an antibody column to preconcentrate the protein and an integrated tip for PS ionization. The 3D printed cartridge drastically improves the ability to detect proteins, including post translational modifications, using PS-MS. In the final project, paper-based surface enhanced Raman spectroscopy (pSERS) was coupled with PS-MS using a 3D printed insert that could facilitate both analytical techniques from the same sample substrate. In this work, organophosphate molecules were analyzed using this dual-technique assay, with a total analysis time of less than five minutes. The three examples discussed are a small subsection of PS-MS projects that benefited from the use of 3D printing.

Presentation 4
3D Printing and Bioprinting as a Rapid Prototyping Approach for Applications in Cell Biology and Tissue Engineering
Eric Spivey, Vanderbilt University
3D printing and other methods of additive manufacturing have improved rapidly in recent years, and have proven especially valuable in the areas of bespoke manufacturing, distributed manufacturing and rapid prototyping. 3D bioprinting, a specialized form of 3D printing which incorporates biomaterials and living cells as “bioink”, has improved in tandem with these more general methods. In particular, 3D bioprinters now print a broad assortment of thermal-setting, shear-thinning and photocrosslinking hydrogels with spatial features that range from submicron to centimeter scale. This presentation will focus on how 3D printing and bioprinting enable new analytical opportunities in tissue engineering and cell biology, via the rapid prototyping of complex high-spatial resolution structures, and biological structures with defined functional features. Specific examples will highlight the use of 3D printing to provide rapid prototyping for organ-on-chip and microdissector devices, and attempts to make the biological components produced in these devices more accessible to analysis.

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