Molecular, Cellular, and Tissue Engineering Commons^™

High-Throughput Robotic Ethanol Inhibition Assays For Engineered Thermophilic Biofuel Strains, Kevin He, Daniel Olson, Marybeth Maloney, Anthony Lanahan

Wetterhahn Science Symposium Posters

Ethanol stress assays are commonly used to evaluate microbial tolerance, metabolic adaptation, and fermentation performance. However, manual liquid handling introduces variability across replicate wells and small-volume pipetting steps, limiting reproducibility and throughput. This study developed an automated OT-2 robotic workflow to generate replicated ethanol concentration gradients for high-throughput inhibition assays in engineered thermophilic biofuel strains. Kinetic plate-reader measurements were used to quantify ethanol-dependent growth responses under anaerobic fermentation conditions. The reasearch question is: How do engineered thermophilic biofuel strains differ in ethanol-dependent growth inhibition under anaerobic fermentation conditions, and can automated robotic assays improve the reproducibility of these measurements? Can …

Quantitative Fluorescence Imaging Using A Paired Agent Approach For Improving Surgical Decision Making, Sanjana Pannem

Dartmouth College Ph.D Dissertations

Surgical resection remains to be the cornerstone of treatment for most solid tumors, where the primary objective is to remove all visible and microscopic disease. However, the extent of tumor resection is inherently limited by the surgeon’s ability to distinguish tumor from normal tissue, intraoperatively. As a result, positive surgical margins (PSM), where residual tumor remains after surgery, are a common occurrence and are associated with increased recurrence, need for adjuvant therapies, dismal patient outcomes, and high healthcare costs.

Fluorescence guided surgery (FGS) has emerged as a promising technique to enhance intraoperative visualization of tumors and may help reduce the …

Advancing Computational Analysis Of Fluorescence Imaging For Head And Neck Cancer Surgical Guidance, Yao Chen

Dartmouth College Ph.D Dissertations

The main goal of surgery for head and neck squamous cell carcinoma (HNSCC) is to remove the tumor completely while preserving as much healthy tissue as possible to enhance patient survival and reduce complications. However, achieving clear surgical margins is difficult due to the complex anatomy of the head and neck and the reliance on a surgeon’s visual and tactile assessments. Recently, near-infrared (NIR) fluorescence molecular imaging has shown potential for better tumor visualization during surgery, using targeted imaging agents like epidermal growth factor receptor (EGFR) probes. This has led to the development of fluorescence-guided surgery (FGS), which helps surgeons …

Design And Validation Of A Novel Patterned Microfluidic Culture Platform For Hipsc-Cm Maturation, Anisia Gabriela Țiplea

Dartmouth College Master’s Theses

Adult cardiomyocytes (CMs) are among the most challenging cells to culture in vitro, given their poor adherence to plastic culture wells and poor maturation outside the native extracellular matrix (Narkar et al., 2022). Human induced pluripotent stem cell-derived CMs (hiPSC-CMs) offer a viable alternative to study heart development and disease progression. While not comparable to adult CMs in their maturity (Narkar et al., 2022), they mimic the clinical phenotypes of heart failure and are more feasible to culture. Nevertheless, hiPSC-CMs require specialized culture conditions that cannot be achieved with 2D static culturing, which fail to replicate native tissue environments (X. …

Characterization Of Influenza-Specific Antibodies, Xinping Ran

ENGS 86 Independent Projects (AB Students)

Influenza viruses cause respiratory illness that affects 8% of the U.S. population annually. However, current influenza vaccines have a limited efficacy of only 40-60%. This highlights the need for a universal vaccine design that can generate broadly neutralizing antibodies against influenza surface protein hemagglutinin (HA). Previous work by our group identified HA-specific monoclonal antibodies (mAbs) from serum samples through high-throughput antibody profiling. In this study, three of the identified mAbs were generated using mammalian cell expression and their binding affinity against three HA antigens was characterized via Enzyme-Linked Immuno-Sorbent Assay (ELISA). The ELISA results demonstrated specific binding affinity towards B/Yamagata/18/1998 …

Multimodal Ultrasound Imaging For Improved Metastatic Lymph Node Detection, Sidhartha Jandhyala

Dartmouth College Ph.D Dissertations

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide and is complex in nature due to the variety of organs located in the head and neck region. Knowing the metastatic state of the lymph nodes is paramount in accurately staging and treating HNSCC patients. Currently, metastatic lymph node detection involves the use of magnetic resonance imaging and/or x-ray computed tomography, followed by biopsies for histological confirmation. The main diagnostic criteria is the size of the nodes; however, current imaging methods are not 100% accurate due natural lymph node variability. Ultrasound imaging is able to provide …

Bio Circuits For Evolutionary Biotech, Camilo Toruno

ENGS 86 Independent Projects (AB Students)

The field of bioengineering has much promise for renewable chemical production, bioremediation, and of course medical applications. Developing new useful microorganisms is extremely time and capital intensive, typically taking 50 million USD and eight years. This is due in large part to the low throughput techniques that are characteristic of the field of metabolic engineering. Here we describe the modification of an existing synthetic biosensor to measure the pharmaceutical dopamine, and the use of a circuit simulator Cadence to predict improvements to the biosensor. This biosensor paired with directed evolution techniques could reach throughputs of 5 million cells per day …

Developing A Control System To Better Understand The Effects Of Pyruvate Decarboxylase Activity On Clostridium Thermocellum Metabolism, Nicholas Cervenka

ENGS 88 Honors Thesis (AB Students)

In order for cellulosic biofuels from Clostridium thermocellum to be commercially viable, the ethanol yield and titer of the microbe must be increased. To accomplish this, it has been suggested to introduce the Pyruvate Decarboxylase (PDC) enzyme into C. thermocellum. In order to demonstrate effects on ethanol production by PDC prior to genetic modification, a cell free system (CFS) has been developed. A purified enzyme system was developed with the CFS to function as a control. Using the purified enzyme system, PDC from Saccharomyces cerevisiae was demonstrated to be a good candidate for further testing in the CFS.