SLAS Technology Authors Talk Tech

Dave Pechter, M.S.M.E.

We invite you to "get behind the science" with SLAS Technology Editorial Board Member and Podcast Editor Dave Pechter, M.S.M.E. (PerkinElmer, Cambridge, MA) and hear from our featured SLAS Technology authors! This podcast series is a chance for readers to meet the people behind the journal science and hear directly from them about their work, their motivations, as well as the context and potential impact of their work. Watch for a new featured author interview with each published issue! read less
ScienceScience

Episodes

Traceable Impedance-Based Dispensing and Cloning of Living Single Cells and Impedance-Based Single-Cell Pipetting
Jun 1 2020
Traceable Impedance-Based Dispensing and Cloning of Living Single Cells and Impedance-Based Single-Cell Pipetting
Volume 25 Issue 3, June 2020Dave Pechter discusses with Georges Muller & Yann Barrandon their two featured research articles, "Traceable Impedance-Based Dispensing and Cloning of Living Single Cells" and "Impedance-Based Single-Cell Pipetting."Traceable Impedance-Based Dispensing and Cloning of Living Single Cells: Single-cell cloning is essential in stem cell biology, cancer research, and biotechnology. Regulatory agencies now require an indisputable proof of clonality that current technologies do not readily provide. Here, we report a one-step cloning method using an engineered pipet combined with an impedance-based sensing tip. This technology permits the efficient and traceable isolation of living cells, stem cells, and cancer stem cells that can be individually expanded in culture and transplanted.Impedance-Based Single-Cell Pipetting: Many biological methods are based on single-cell isolation. In single-cell line development, the gold standard involves the dilution of cells by means of a pipet. This process is time-consuming as it is repeated over several weeks to ensure clonality. Here, we report the modeling, designing, and testing of a disposable pipet tip integrating a cell sensor based on the Coulter principle. We investigate, test, and discuss the effects of design parameters on the sensor performances with an analytical model. We also describe a system that enables the dispensing of single cells using an instrumented pipet coupled with the sensing tip. Most importantly, this system allows the recording of an impedance trace to be used as proof of single-cell isolation. We assess the performances of the system with beads and cells. Finally, we show that the electrical detection has no effect on cell viability.
CURATE.AI: Optimizing Personalized Medicine with Artificial Intelligence
Apr 1 2020
CURATE.AI: Optimizing Personalized Medicine with Artificial Intelligence
Volume 25 Issue 2, April 2020Dave Pechter discusses with Agata Blasiak & Theodore Kee regarding their article, "CURATE.AI: Optimizing Personalized Medicine with Artificial Intelligence." The clinical team attending to a patient upon a diagnosis is faced with two main questions: what treatment, and at what dose? Clinical trials’ results provide the basis for guidance and support for official protocols that clinicians use to base their decisions upon. However, individuals rarely demonstrate the reported response from relevant clinical trials, often the average from a group representing a population or subpopulation. The decision complexity increases with combination treatments where drugs administered together can interact with each other, which is often the case. Additionally, the individual’s response to the treatment varies over time with the changes in his or her condition, whether via the indication or physiology. In practice, the drug and the dose selection depend greatly on the medical protocol of the healthcare provider and the medical team’s experience. As such, the results are inherently varied and often suboptimal. Big data approaches have emerged as an excellent decision-making support tool, but their application is limited by multiple challenges, the main one being the availability of sufficiently big datasets with good quality, representative information. An alternative approach—phenotypic personalized medicine (PPM)—finds an appropriate drug combination (quadratic phenotypic optimization platform [QPOP]) and an appropriate dosing strategy over time (CURATE.AI) based on small data collected exclusively from the treated individual. PPM-based approaches have demonstrated superior results over the current standard of care. The side effects are limited while the desired output is maximized, which directly translates into improving the length and quality of individuals’ lives.
Automated System for Small-Population Single-Particle Processing Enabled by Exclusive Liquid Repellency
Dec 1 2019
Automated System for Small-Population Single-Particle Processing Enabled by Exclusive Liquid Repellency
Volume 24 Issue 6, December 2019Dave Pechter discusses with Chao Li the article, "Automated System for Small-Population Single-Particle Processing Enabled by Exclusive Liquid Repellency." Lossless processing and culture of rare cells (e.g., circulating tumor cells, drug-persistent microorganisms) at single-cell level is of great significance in understanding the heterogeneity of carcinogenesis or human pathogenesis caused by microbial infection. Current single-cell isolation techniques like fluorescence-activated cell sorting (FACS) require relatively large sample volume and cell number to work with, and inflict sample loss and reduced cell viability from the processing. While microfluidic and single-cell printing techniques allow the handling of minute amounts of cellular samples, they either come with limited physical access to the sample of interest due to the closed-channel design (e.g., droplet microfluidics) or sample loss during aspiration, transfer, and sample retrieval from culture.Recently, we reported an extreme wettability phenomenon, named exclusive liquid repellency or ELR. ELR is observed in solid-liquid-liquid three phase systems, where a solid surface shows complete repellency to a liquid (with Young’s contact angle, CA = 180o) when exposed to a second immiscible liquid. This phenomenon is observed when a particular thermodynamic boundary condition is satisfied (i.e., γS/Lcp + γLdp/Lcp ≤ γS/Ldp, where γ - interfacial tension, S - solid, Lcp - liquid of continuous phase, and Ldp - liquid of dispersed phase). Neither surfactant nor flow condition is required, e.g., compared with droplet microfluidics. ELR enables additional fluidic control, robust on-chip cell culture, and improved processing of rare cell samples in open aqueous fluid under oil. ELR is distinct from other liquid repellent systems with CA In this work, we developed an automated platform using ELR microdrops for lossless single-particle (or single-cell) isolation, identification, and retrieval. It features the combined use of a robotic liquid handler, an automated microscopic imaging system, and real-time image-processing software for single-particle identification. The automated ELR technique enables rapid, hands-free, and robust isolation of microdrop-encapsulated rare cellular samples, and further on-chip cell culture or down-stream analysis (e.g., RNA extraction and RT-qPCR).
Technologies for the Directed Evolution of Cell Therapies
Aug 1 2019
Technologies for the Directed Evolution of Cell Therapies
Volume 24 Issue 4, August 2019UCLA Professor, Dino Di Carlo, discusses his review paper, "Technologies for the Directed Evolution of Cell Therapies." The next generation of therapies is moving beyond the use of small molecules and proteins to using whole cells. Compared with the interactions of small-molecule drugs with biomolecules, which can largely be understood through chemistry, cell therapies act in a chemical and physical world and can actively adapt to that world, amplifying complexity but also the potential for truly breakthrough impact. Although there has been success in introducing targeting proteins into cells to achieve a therapeutic effect, for example, chimeric antigen receptor (CAR) T cells, our ability to engineer cells is generally limited to introducing proteins, but not modulating large-scale traits or structures of cellular “machines,” which play critical roles in disease. Example traits include the ability to secrete compounds, deform through tissue, adhere to surrounding cells, apply force to phagocytose targets, or move through extracellular matrix. There is an opportunity to increase the efficacy of cell therapies through the use of quantitative automation tools, to analyze, sort, and select rare cells with beneficial traits. Combined with methods of genetic or epigenetic mutagenesis to create diversity, such approaches can enable the directed cellular evolution of new therapeutically optimal populations of cells and uncover genetic underpinnings of these optimal traits.