Microfluidic chip for spatially and temporally controlled biochemical gradient generation in standard cell-culture Petri dishes

Sahai, Ranjana; Cecchini, Marco; Klingauf, Mirko; Ferrari, Aldo; Martino, Chiara; Castrataro, Piero; Lionetti, Vincenzo; Menciassi, Arianna; Beltram, Fabio

doi:10.1007/s10404-011-0841-2

Cited by 8 publications

(5 citation statements)

References 33 publications

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“…PDMS microfluidic chips, created through conventional microfabrication processes, have been bonded to the Petri dish using O 2 plasma bonding. However, it is important to note that the Petri dish functions not as the central element of the microfluidic chip but rather as a practical vessel for chip operation and observation. , These devices also require a protein coating to facilitate cell adhesion. In certain investigations, researchers have achieved cell culture on PDMS covers bonded to Petri dish substrates, omitting the protein coating step.…”

Section: Introductionmentioning

confidence: 99%

Composite Microfluidic Petri Dish-Chip (MPD-Chip) without Protein Coating for 2D Cell Culture

Yin,

Lu,

Liu

et al. 2023

Langmuir

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Hydrophilicity is a requisite attribute for the 2D cell culture substrate's surface, facilitating cell adhesion and spreading. Conventional poly(dimethylsiloxane) (PDMS) microfluidic chips necessitate protein coatings to enhance hydrophilicity; however, this approach is afflicted by issues of transient efficacy, interference with cell analysis, and high costs. This paper presents a protein-free microfluidic chip, termed a "microfluidic Petri dish-chip (MPD-chip)", integrating PDMS as the cover and a tissue culture-treated (TC-treated) Petri dish as the substrate. Microstructures are hot-embossed onto the Petri dish substrate using a silicon mold. This meticulous replication process serves to establish stable flow field dynamics within the chip. A simplified method for irreversible bonding, utilizing plasma activation and silylation, is proposed for affixing the PDMS cover onto the microstructured Petri dish substrate. The prepared composite chip exhibits remarkable tightness, boasting a notable bond strength of 2825 kPa. Furthermore, the composite microfluidic chip demonstrates the capability to withstand flow velocities of at least 200 μL/min, effectively meeting the required injection standards for both cell suspension and culture medium. SH-SY5Y and HeLa cells are cultured dynamically in the MPD-chip and control groups. Outcomes encompassing normalized cell density, cell adhesion area, and cell viability metrics unequivocally highlight the superiority of the MPD-chip in facilitating long-term two-dimensional (2D) cell cultures.

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Section: Introductionmentioning

confidence: 99%

Composite Microfluidic Petri Dish-Chip (MPD-Chip) without Protein Coating for 2D Cell Culture

Yin,

Lu,

Liu

et al. 2023

Langmuir

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“…This was achieved by exploiting some characteristics of the microuidic regime, such as the laminar ow. 10,11 A number of biodevices were proposed to study relevant cell biology aspects, such as cell migration, 12,13 axon guidance 14 and stem cell differentiation. 15 Although devices such as these did show great potential, several drawbacks have limited their widespread use in cell biology.…”

Section: Introductionmentioning

confidence: 99%

Tubeless biochip for chemical stimulation of cells in closed-bioreactors: anti-cancer activity of the catechin–dextran conjugate

et al. 2014

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The most exciting promise of cell micro-bioreactors is their expected ability to reproduce specific aspects of physiological environments in vitro, singularly or in combination, and to perform advanced biological studies to unravel novel interaction mechanisms; but this challenge is formidable. Moreover, the use of microfluidics is limited by the difficulty to adapt standard tissue culture protocols to miniaturised biochips and by the need of bulky external equipment such as liquid pumping systems. Here, we introduce an original microfluidic gradient generator for culturing cells in closed microchambers, based on: (1) automatic cell valving, (2) hydrostatic-pressure pumping, and (3) on-chip liquid reservoirs. The biochip is designed to be fully compatible with standard biological procedures and to operate with no external control units or cumbersome ancillary components. Remarkably, the use of integrated liquid reservoirs prevents air bubble formation and associated channel clogging. We detail the rationale behind the fluidic design, simulate and measure the chip fluid-dynamics by finite element simulations and microflow velocimetry, respectively. We finally exploit our chip to study the kinetics of an anti-cancer molecule, the catechin-dextran conjugate, on HeLa cells, demonstrating a very fast action. We argue that the presented fluidic concept and architecture are not limited to the present realization, but can be easily applied to other chip geometries and adapted to a number of biological studies

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“…At the heart of this success is the capability to control the properties of fluid-velocity fields at the microscale allowing a precise spatial and temporal modulation of chemical-concentration , and shear-stress profiles. − Hence, development of rapid and easy methods to measure fluid-velocity fields prior or during biological experiments is of paramount importance. Several techniques were developed in the past decade to measure the velocity field inside microfluidic devices for both fundamental fluid-dynamics studies and device testing.…”

mentioning

confidence: 99%

“…1,2 Among these, biology is probably the field of choice for LOC applications: several traditional assays have already been miniaturized within micrototal analysis systems (μTAS); 3 high-throughput devices were designed for parallel biomolecule screening, 4 and a variety of biomimetic LOC devices were realized to study complex cellular cultures. 5,6 At the heart of this success is the capability to control the properties of fluid-velocity fields at the microscale allowing a precise spatial and temporal modulation of chemical-concentration 7,8 and shear-stress profiles. 9−11 Hence, development of rapid and easy methods to measure fluid-velocity fields prior or during biological experiments is of paramount importance.…”

mentioning

confidence: 99%

Easy Monitoring of Velocity Fields in Microfluidic Devices Using Spatiotemporal Image Correlation Spectroscopy

et al. 2013

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Spatiotemporal image correlation spectroscopy (STICS) is a simple and powerful technique, well established as a tool to probe protein dynamics in cells. Recently, its potential as a tool to map velocity fields in lab-on-a-chip systems was discussed. However, the lack of studies on its performance has prevented its use for microfluidics applications. Here, we systematically and quantitatively explore STICS microvelocimetry in microfluidic devices. We exploit a simple experimental setup, based on a standard bright-field inverted microscope (no fluorescence required) and a high-fps camera, and apply STICS to map liquid flow in polydimethylsiloxane (PDMS) microchannels. Our data demonstrates optimal 2D velocimetry up to 10 mm/s flow and spatial resolution down to 5 μm.

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Microfluidic chip for spatially and temporally controlled biochemical gradient generation in standard cell-culture Petri dishes

Cited by 8 publications

References 33 publications

Composite Microfluidic Petri Dish-Chip (MPD-Chip) without Protein Coating for 2D Cell Culture

Composite Microfluidic Petri Dish-Chip (MPD-Chip) without Protein Coating for 2D Cell Culture

Tubeless biochip for chemical stimulation of cells in closed-bioreactors: anti-cancer activity of the catechin–dextran conjugate

Easy Monitoring of Velocity Fields in Microfluidic Devices Using Spatiotemporal Image Correlation Spectroscopy

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