Laser-fabricated cell patterning stencil for single cell analysis

Messner, Jacob; Glenn, Honor L.; Meldrum, Deirdre R.

doi:10.1186/s12896-017-0408-8

Cited by 10 publications

(6 citation statements)

References 77 publications

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“…A different approach that allows both to increase complexity within conventional culture vessels and to maintain an open well system are stencil-like plating devices [ 40 , 41 ]. These systems are temporary structures that guide the organisation of cells within an open well, although at present they suffer from the same fabrication limitations as the abovementioned strategies.…”

Section: Resultsmentioning

confidence: 99%

Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

Hagemann,

Bailey,

Carraro

et al. 2024

PLoS Biol

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Cell culture devices, such as microwells and microfluidic chips, are designed to increase the complexity of cell-based models while retaining control over culture conditions and have become indispensable platforms for biological systems modelling. From microtopography, microwells, plating devices, and microfluidic systems to larger constructs such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques, equipment and tools required for their manufacture, and the expertise necessary, biological and biomedical labs tend more often to rely on already made devices. Indeed, commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customisation by each individual lab. The last point is quite crucial, as often experiments in wet labs are adapted to whichever design is already available rather than designing and fabricating custom systems that perfectly fit the biological question. This combination of factors still restricts widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent advances in bioengineering and microfabrication aimed at solving these issues, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline. This is thought specifically for biomedical and biological wet labs with not prior experience in the field, which will enable them to generate a wide variety of customisable devices for cell culture and tissue engineering in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with limited expertise in those techniques and enables the manufacture of complex devices across the μm to cm scale. We provide a ready-to-go pipeline for the efficient treatment of resin-based 3D-printed constructs for PDMS curing, using a combination of polymerisation steps, washes, and surface treatments. Together with the extensive characterisation of the fabrication pipeline, we show the utilisation of this system to a variety of applications and use cases relevant to biological experiments, ranging from micro topographies for cell alignments to complex multipart hydrogel culturing systems. This methodology can be easily adopted by any wet lab, irrespective of prior expertise or resource availability and will enable the wide adoption of tailored microfabricated devices across many fields of biology.

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Section: Resultsmentioning

confidence: 99%

Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

Hagemann,

Bailey,

Carraro

et al. 2024

PLoS Biol

View full text Add to dashboard Cite

show abstract

“…parylene 43 ) or extensive time and resources to fabricate, which makes them difficult to implement as a routine system for most biology labs. Another approach that allows increased levels of complexity within conventional culture vessels and maintaining an open well system are stencil-like devices 44, 45 , although at present these also suffer the same technical limitations as the above-mentioned strategies. Moreover, this method is especially affected by the geometrical limitations of feature extrusion across volumes resulting in thin devices with limited possibility of customisation.…”

Section: Resultsmentioning

confidence: 99%

“…A different approach that allows both to increase complexity within conventional culture vessels and to maintain an open well system are stencil-like plating devices 33, 34 . These systems are temporary structures that guide the organisation of cells within an open well, although at present they suffer from the same fabrication limitations as the above-mentioned strategies.…”

Section: Resultsmentioning

confidence: 99%

SOL3D: Soft-lithography on 3D vat polymerised moulds for fast, versatile, and accessible high-resolution fabrication of customised multiscale cell culture devices with complex designs

Hagemann

Bailey

Khokhar

et al. 2022

Preprint

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Commercially available cell culture devices are designed to increase the complexity of simple cell culture models to provide better experimental platforms for biological systems. From microtopography, microwells, plating devices and microfluidic systems to larger constructs for specific applications like live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology labs. However, the techniques used for their fabrication can be out of reach for most wet labs due to cost and availability of specialised equipment or the need for engineering expertise. Moreover, these techniques also have technical limitations to the volumes, shapes and dimensions they can generate. For these reasons, creating customisable devices tailored to lab-specific biological questions remains difficult to apply. Taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft-lithography we have developed an optimised microfabrication pipeline capable of generating a wide variety of customisable devices for cell culture and tissue engineering in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This technique enables the manufacture of complex devices across scales bridging the gap between microfabrication and fused deposition moulding (FDM) printing. The method we describe allows for the efficient treatment of resin-based 3D printed constructs for PDMS curing, using a combination of curing steps, washes and surface treatments. Together with the extensive characterisation of the fabrication pipeline, we provide several proof-of-principle applications ranging from simple 2D culture devices to large tissue engineering constructs and organoid formation systems. We believe this methodology will be applicable in any wet lab, irrespective of prior expertise or resource availability and will therefore enable a wide adoption of tailored microfabricated devices across many fields of biology.

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“…In addition to the stamp method, another promising method is to study cell localization directly using the structure of topographic patterns (physical modications, such as micropits and nanowires). [20][21][22] The substrate used in this method is generally silicon, so there is no shelf-life problem, and it is easy to obtain complex topographical patterns, but its localization efficiency needs to be further improved. Therefore, developing robust topographical patterns to improve the localization ratio of single-cells is still a challenging issue.…”

Section: Introductionmentioning

confidence: 99%