Mapping of Enzyme Kinetics on a Microfluidic Device

Rho, Hoon Suk; Hanke, Alexander T.; Ottens, Marcel; Gardeniers, Han

doi:10.1371/journal.pone.0153437

Cited by 19 publications

(20 citation statements)

References 37 publications

(33 reference statements)

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“…The resulting data (Figure f) were used for the calculation of reaction rates that were plotted against the substrate concentration (Figure g). In case of the SLA Chip the V0/[S] graph showed the expected plateau toward higher substrate concentrations, thus leading to an estimated Michaelis‐Menten constant (Km) of 31 × 10 ‐6 m , which is lower than the Km values obtained from microplate experiments (Km ≈ 100–200 × 10 ‐6 m ) . Interestingly, the reaction in the smaller MM droplets occurred substantially faster, as clearly indicated by the truncation of the initial phase of signal development (Figure f) as well as by the linear slope of the V0/[S] plot, which does not reach the expected plateau within the concentration range employed.…”

Section: Resultsmentioning

confidence: 92%

Microfluidic Chips for Life Sciences—A Comparison of Low Entry Manufacturing Technologies

Grösche

Zoheir

Stegmaier

et al. 2019

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mixing. These features can often accelerate or even enable reactions to proceed. [4] A key factor for the successful dissemination of this technology is the ability for production of masters for molding microfluidic droplet generators and related accessories through rapid prototyping with low associated manufacturing and material costs, because only few ready-to-use microfluidic devices are currently commercially available and their geometries are usually limited to standard applications.Soft lithography using SU-8 masters is the gold standard for the production of microfluidic prototypes. [5] In this method, an SU-8 photoresist is structured by means of optical lithography and the resulting negative structure is then molded by soft lithography into polydimethylsiloxane (PDMS). For sealing, the elastomeric PDMS chips are usually plasma bonded onto glass supports. The very high structural resolution along with excellent surface quality in terms of low roughness is the major advantages of this technology, whereas low aspect ratios and difficulties in the production of variable heights in a single chip, along with high machine costs and the requirement of clean room facilities for the production of masters, are on the downside. Since trained clean room personnel and expensive infrastructure are often not available in research institutes engaged in biomedical research and the life sciences, there is a great demand for alternative costeffective manufacturing processes for microfluidic chips.The implementation of additive manufacturing (AM) methods in microfluidic prototyping is currently attracting Microfluidic water-in-oil droplets are a versatile tool for biological and biochemical applications due to the advantages of extremely small monodisperse reaction vessels in the pL-nL range. A key factor for the successful dissemination of this technology to life science laboratory users is the ability to produce microfluidic droplet generators and related accessories by low-entry barrier methods, which enable rapid prototyping and manufacturing of devices with low instrument and material costs. The direct, experimental side-by-side comparison of three commonly used additive manufacturing (AM) methods, namely fused deposition modeling (FDM), inkjet printing (InkJ), and stereolithography (SLA), is reported. As a benchmark, micromilling (MM) is used as an established method. To demonstrate which of these methods can be easily applied by the non-expert to realize applications in topical fields of biochemistry and microbiology, the methods are evaluated with regard to their limits for the minimum structure resolution in all three spatial directions. The suitability of functional SLA and MM chips to replace classic SU-8 prototypes is demonstrated on the basis of representative application cases. Microfluidic ManufacturingProf. R. Mikut

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Section: Resultsmentioning

confidence: 92%

Microfluidic Chips for Life Sciences—A Comparison of Low Entry Manufacturing Technologies

Grösche

Zoheir

Stegmaier

et al. 2019

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“…Microfluidic devices were fabricated using polydimethylsiloxane (PDMS) multilayer softlithography and we followed the protocols used in our previous studies . After the final adhesion step, the device was autoclaved for 20 min at 120°C for sterilization.…”

Section: Methodsmentioning

confidence: 99%

Parallel probing of drug uptake of single cancer cells on a microfluidic device

et al. 2017

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Drug resistance is frequently developing during treatment of cancer patients. Intracellular drug uptake is one of the important characteristics to understand mechanism of drug resistance. However, the heterogeneity of cancer cells requires the investigation of drug uptake at the single cell level. Here, we developed a microfluidic device for parallel probing of drug uptake. We combined a v-type valve and peristaltic pumping to select individual cells from a pool of prostate cancer cells (PC3) and place them successively in separate cell chambers in which they were exposed to the drug. Six different concentrations of doxorubicin, a naturally fluorescent anti-cancer drug, were created in loop-shaped reactors and exposed to the cell in closed 2 nL volume chambers. Monitoring every single cell over time in 18 parallel chambers revealed increased intracellular fluorescence intensity according to the dose of doxorubicin, as well as nuclear localization of the fluorescent drug after 2 h of incubation. The herein proposed technology demonstrated a first series of proof of concept experiments and it shows high potential to use for probing drug sensitivity of single cancer cell.

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“…Niu et al (2011) developed a droplet-based platform capable of performing dilutions within a range of four orders of magnitude by splitting and (re)merging droplets to create reagent gradients (Niu et al, 2011). Rho et al (2016) used peristaltic mixing in controlled volume microreactors to generate stepwise concentration gradients of two reagents (Rho et al, 2016). The induction of…”

Section: Available Unit Operations In Microfluidic Chipsmentioning

confidence: 99%

“…Combination or blending of two or more substances or compounds  Passive mixing, such as channel topology (Liau et al, 2005), contact area increase (Lee et al, 2011), lamination (Gürsel et al, 2017), coiled flow inverter (Klutz et al, 2015) or stimuli-responsive hydrogels (Prettyman and Eddington, 2011);  Active mixing, such as acoustically-induced microstreams, dielectrophoretic micromixers, electrokinetic actuation (Gao and Gui, 2016), velocity pulsing and magneto-hydrodynamic flow have also been thoroughly developed and applied (Lee et al, 2011) Dilutions Definition of gradients of a target substance  Droplet-based (Niu et al, 2011);  Peristaltic mixing (Rho et al, 2016);…”

Section: Mixingmentioning

confidence: 99%

“Connecting worlds – a view on microfluidics for a wider application”

Fernandes

Gernaey

Krühne

2018

Biotechnology Advances

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From its birth, microfluidics has been referenced as a revolutionary technology and the solution to long standing technological and sociological issues, such as detection of dilute compounds and personalized healthcare. Microfluidics has for example been envisioned as: (1) being capable of miniaturizing industrial production plants, thereby increasing their automation and operational safety at low cost; (2) being able to identify rare diseases by running bioanalytics directly on the patient's skin; (3) allowing health diagnostics in point-of-care sites through cheap lab-on-a-chip devices. However, the current state of microfluidics, although technologically advanced, has so far failed to reach the originally promised widespread use. In this paper, some of the aspects are identified and discussed that have prevented microfluidics from reaching its full potential, especially in the chemical engineering and biotechnology fields, focusing mainly on the specialization on a single target of most microfluidic devices and offering a perspective on the alternate, multi-use, "plug and play" approach. Increasing the flexibility of microfluidic platforms, by increasing their compatibility with different substrates, reactions and operation conditions, and other microfluidic systems is indeed of surmount importance and current academic and industrial approaches to modular microfluidics are presented. Furthermore, two views on the commercialization of plug-and-play microfluidics systems, leading towards improved acceptance and more widespread use, are introduced. A brief review of the main materials and fabrication strategies used in these fields, is also presented. Finally, a step-wise guide towards the development of microfluidic systems is introduced with special focus on the integration of sensors in microfluidics. The proposed guidelines are then applied for the development of two different example platforms, and to three examples taken from literature. With this work, we aim to provide an interesting perspective on the field of microfluidics when applied to chemical engineering and biotechnology studies, as well as to contribute with potential solutions to some of its current challenges.

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Mapping of Enzyme Kinetics on a Microfluidic Device

Cited by 19 publications

References 37 publications

Microfluidic Chips for Life Sciences—A Comparison of Low Entry Manufacturing Technologies

Microfluidic Chips for Life Sciences—A Comparison of Low Entry Manufacturing Technologies

Parallel probing of drug uptake of single cancer cells on a microfluidic device

“Connecting worlds – a view on microfluidics for a wider application”

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