Femtosecond laser machined microfluidic devices for imaging of cells during chemotaxis

Costa, Lino; Terekhov, Alexander; Rajput, Deepak; Hofmeister, William; Jowhar, Dawit; Wright, Gus A.; Janetopoulos, Christopher

doi:10.2351/1.3614405

Cited by 11 publications

(11 citation statements)

References 30 publications

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“…For the large horizontal channels in Supplementary Figure 6, toothpicks were used to mold the PDMS channels. All femtosecond machining of glass was performed at UTSI as previously described (Costa et al, 2011; Wright et al, 2012). Micromirrored wells were etched in silicon wafers (Seale et al, 2008), cut, and glued to the glass slide.…”

Section: Methodsmentioning

confidence: 99%

“…Most importantly, we have developed glass bases that have microchannels machined into them. Recent advances in femtosecond laser machining allow us to make three-dimensional (3D) channels in glass (Ke et al, 2005; White et al, 2008; Costa et al, 2011). These fluid channels can permit exchange of fresh medium while the specimen is being studied, and also permit introduction of drugs or other chemicals whose effects on the organism or cell can be observed in real time.…”

Section: Introductionmentioning

confidence: 99%

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A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

et al. 2014

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A microcompressor is a precision mechanical device that flattens and immobilizes living cells and small organisms for optical microscopy, allowing enhanced visualization of sub-cellular structures and organelles. We have developed an easily fabricated device, which can be equipped with microfluidics, permitting the addition of media or chemicals during observation. This device can be used on both upright and inverted microscopes. The apparatus permits micrometer precision flattening for nondestructive immobilization of specimens as small as a bacterium, while also accommodating larger specimens, such as Caenorhabditis elegans, for long-term observations. The compressor mount is removable and allows easy specimen addition and recovery for later observation. Several customized specimen beds can be incorporated into the base. To demonstrate the capabilities of the device, we have imaged numerous cellular events in several protozoan species, in yeast cells, and in Drosophila melanogaster embryos. We have been able to document previously unreported events, and also perform photobleaching experiments, in conjugating Tetrahymena thermophila.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

et al. 2014

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“…The devices were fabricated and assembled in an ISO 1000 class clean room, in accordance with the procedure outlined in detail in Costa et al (2011). The microfluidic channels and the chemical concentration gradient forming structures were patterned on ultraviolet grade fused silica using the femtosecond laser micromachining system (White et al, 2008; Zalloum et al, 2010).…”

Section: Methodsmentioning

confidence: 99%

On-Chip Open Microfluidic Devices for Chemotaxis Studies

et al. 2012

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Microfluidic devices can provide unique control over both the chemoattractant gradient and the migration environment of the cells. Our work incorporates laser-machined micro and nanofluidic channels into bulk fused silica and cover slip-sized silica wafers. We have designed “open” chemotaxis devices that produce passive chemoattractant gradients without an external micropipette system. Since the migration area is unobstructed, cells can be easily loaded and strategically placed into the devices with a standard micropipette. The reusable monolithic glass devices have integral ports that can generate multiple gradients in a single experiment. We also used cover slip microfluidics for chemotaxis assays. Passive gradients elicited from these cover slips could be readily adapted for high throughput chemotaxis assays. We have also demonstrated for the first time that cells can be recruited into cover slip ports eliciting passive chemoattractant gradients. This proves, in principle, that intravital cover slip configurations could deliver controlled amounts of drugs, chemicals, or pathogens as well as recruit cells for proteomic or histological analysis in living animals while under microscopic observation. Intravital cover slip fluidics will create a new paradigm for in vivo observation of biological processes.

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“…27,28,34 The microfluidic device was designed in AutoCAD, laser ablated with a femtosecond laser machining station, and etched in 10 M KOH solution at 80 C for 1 h. The device was fabricated and assembled in an ISO 1000 class clean room at the Center for Laser Application of the University of Tennessee Space Institute. Two mm diameter inlet/outlet and culture-well holes were drilled through 500-lm thick fused silica wafers.…”

Section: A Device Fabrication and Preparationmentioning

confidence: 99%

“…[20][21][22] We have developed a co-culture bioreactor to mimic and examine the breast cancer TME that takes into consideration the need for interstitial flow and the ability to grow numerous cell types in a 3D matrix. [23][24][25][26] This device was machined using femtosecond laser-etching technology 27,28 and mimics the TME and surrounding vasculature. The design incorporates a semicircular chamber connected to a channel 200 lm in width to an array of capillary-like channels, all machined in glass.…”

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confidence: 99%

Engineered three-dimensional microfluidic device for interrogating cell-cell interactions in the tumor microenvironment

et al. 2014

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Stromal cells in the tumor microenvironment play a key role in the metastatic properties of a tumor. It is recognized that cancer-associated fibroblasts (CAFs) and endothelial cells secrete factors capable of influencing tumor cell migration into the blood or lymphatic vessels. We developed a microfluidic device that can be used to image the interactions between stromal cells and tumor cell spheroids in a three dimensional (3D) microenvironment while enabling external control of interstitial flow at an interface, which supports endothelial cells. The apparatus couples a 200-lm channel with a semicircular well to mimic the interface of a blood vessel with the stroma, and the design allows for visualization of the interactions of interstitial flow, endothelial cells, leukocytes, and fibroblasts with the tumor cells. We observed that normal tissue-associated fibroblasts (NAFs) contribute to the "single file" pattern of migration of tumor cells from the spheroid in the 3D microenvironment. In contrast, CAFs induce a rapid dispersion of tumor cells out of the spheroid with migration into the 3D matrix. Moreover, treatment of tumor spheroid cultures with the chemokine CXCL12 mimics the effect of the CAFs, resulting in similar patterns of dispersal of the tumor cells from the spheroid. Conversely, addition of CXCL12 to co-cultures of NAFs with tumor spheroids did not mimic the effects observed with CAF co-cultures, suggesting that NAFs produce factors that stabilize the tumor spheroids to reduce their migration in response to CXCL12.

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Femtosecond laser machined microfluidic devices for imaging of cells during chemotaxis

Cited by 11 publications

References 30 publications

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

On-Chip Open Microfluidic Devices for Chemotaxis Studies

Engineered three-dimensional microfluidic device for interrogating cell-cell interactions in the tumor microenvironment

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