Automated Reagent-Dispensing System for Microfluidic Cell Biology Assays

Ly, Jimmy; Masterman‐Smith, Michael; Ramakrishnan, Ravichandran; Sun, Jing; Kokubun, Brent A.; Dam, R. Michael van

doi:10.1177/2211068213504758

Cited by 8 publications

(8 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In cell biology, most non−steady state experiments are initiated by pipetting a small volume of concentrated solution into the existing culture medium or by completely removing the medium and then replacing it with a medium of different composition, sometimes with an intervening wash. Increasingly, automated microfluidics-based tools are being used to control the composition of the extracellular medium as desired ( Salieb-Beugelaar et al. , 2010 ; Ly et al. , 2013 ; Haque et al.…”

Section: Resultsmentioning

confidence: 99%

Differential equation methods for simulation of GFP kinetics in non–steady state experiments

Phair¹

2018

MBoC

View full text Add to dashboard Cite

Genetically encoded fluorescent proteins, combined with fluorescence microscopy, are widely used in cell biology to collect kinetic data on intracellular trafficking. Methods for extraction of quantitative information from these data are based on the mathematics of diffusion and tracer kinetics. Current methods, although useful and powerful, depend on the assumption that the cellular system being studied is in a steady state, that is, the assumption that all the molecular concentrations and fluxes are constant for the duration of the experiment. Here, we derive new tracer kinetic analytical methods for non–steady state biological systems by constructing mechanistic nonlinear differential equation models of the underlying cell biological processes and linking them to a separate set of differential equations governing the kinetics of the fluorescent tracer. Linking the two sets of equations is based on a new application of the fundamental tracer principle of indistinguishability and, unlike current methods, supports correct dependence of tracer kinetics on cellular dynamics. This approach thus provides a general mathematical framework for applications of GFP fluorescence microscopy (including photobleaching [FRAP, FLIP] and photoactivation to frequently encountered experimental protocols involving physiological or pharmacological perturbations (e.g., growth factors, neurotransmitters, acute knockouts, inhibitors, hormones, cytokines, and metabolites) that initiate mechanistically informative intracellular transients. When a new steady state is achieved, these methods automatically reduce to classical steady state tracer kinetic analysis.

show abstract

Section: Resultsmentioning

confidence: 99%

Differential equation methods for simulation of GFP kinetics in non–steady state experiments

Phair¹

2018

MBoC

View full text Add to dashboard Cite

show abstract

“…Automated cell culture systems enable large-scale production of cells and enhance technical precision, reproducibility and efficiency ( Konagaya et al, 2015 ). Monitoring the flow rate, for example, during media change, is an important operation to ensure that shear forces on cells are contained ( Ly et al, 2013 ). A fine-tuning of the pipetting settings could decrease the shear stress, but very slow aspirating steps are associated with a long duration of the process.…”

Section: Automating Precise Pipettingmentioning

confidence: 99%

Automation, Monitoring, and Standardization of Cell Product Manufacturing

Doulgkeroglou

Nubila

Nießing

et al. 2020

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Although regenerative medicine products are at the forefront of scientific research, technological innovation, and clinical translation, their reproducibility and large-scale production are compromised by automation, monitoring, and standardization issues. To overcome these limitations, new technologies at software (e.g., algorithms and artificial intelligence models, combined with imaging software and machine learning techniques) and hardware (e.g., automated liquid handling, automated cell expansion bioreactor systems, automated colony-forming unit counting and characterization units, and scalable cell culture plates) level are under intense investigation. Automation, monitoring and standardization should be considered at the early stages of the developmental cycle of cell products to deliver more robust and effective therapies and treatment plans to the bedside, reducing healthcare expenditure and improving services and patient care.

show abstract

“…Opened by an extensive review article by Young 1 on microfluidic platforms for studying angiogenesis, a total of 10 articles cover reviews of the latest microscale tools and technologies for in vitro cell culture and analysis [2][3][4][5][6] and original research on development of new automated cell culture platforms and cell patterning and assembly techniques. [7][8][9][10] In the February 2014 issue (volume 19, issue 1), we will publish an exciting collection of articles, particularly in tools and technology associated with measurement, analyses, and delivery. Two comprehensive reviews, one by Meacham et al and the other by Pereira et al, on delivery technology at different length scales, will open this issue.…”

Section: From the Guest Editorsmentioning

confidence: 99%