polymer microfluidic devices  (Biosite Inc)

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Addressing Patient Specificity in the Engineering of Tumor Models

doi: 10.3389/fbioe.2019.00217

Figure Lengend Snippet: Overview of microfluidic-based tumor models using patient-derived materials.

Article Snippet: The spheroid fraction was mixed with collagen-I and used for culture in a cyclic olefin co-polymer (COC)-based 3D microfluidic device (DAX-1, AIM BIOTECH) (Aref et al., ).

Techniques: Chemotaxis Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Addressing Patient Specificity in the Engineering of Tumor Models

doi: 10.3389/fbioe.2019.00217

Figure Lengend Snippet: Microfluidic systems using patient-derived materials for chemotherapy (A–G) and immunotherapy (H,I) . (A) Top: schematic of a patient-derived tumor explant, microdissected, loaded in a microfluidic device and injected with various drugs. Results are then interpreted to identify non-responders to treatment for a personalized treatment strategy. Bottom: photograph of tumor prior and after microdissection in 500 μm-PDMEs and design of the microfluidic device containing up to 5 chambers for PDME entrapment and drug testing. (B) PDMEs from various patients and different cancers are analyzed by live/dead cell assay using confocal microscopy (green = live, red = dead), showing various PDME architectures and viability patterns. (C) PDMEs from an ovarian tumor treated with carboplatin shows high heterogeneity in PDME structure and viability response for PDME originating from the same tumor. (D) Corresponding graphs of viability data from (B) (top) and (C) (bottom) compared to non-treated controls show inter-cancer tumor heterogeneity and drug efficacy on PDMEs from one tumor, despite intra-tumor heterogeneity observed in (C) . (E) Microfluidic device enabling in situ spheroid patterning technique: a microfluidic chamber (i) is filled with a mixture (blue) containing hydrogel precursors, photoinitiator, and patient-derived tumor cells (ii) and then illuminated with UV light through a photomask (gray) (iii). Exposed precursor is crosslinked into a hydrogel (dark blue), detaining cells within the region (iv), and non-crosslinked gel is flushed form the chamber with clean saline from the chamber (v). Finally, saline is replaced with media (red) (vi) for incubation. (F) Schematic of system that offers low volumes and closed loop fluidic circuit controlled by a computer-controlled peristatic pump to treated individual organoids. (G) In vitro chemotherapy assessment of organoids derived from two patients with mesothelioma treated with various drug doses and combinations shows different response between patients, although treatment occurred at different times between patients (viability measured from live/dead confocal microscopy on organoids). (H) Schematic of PDMEs obtained after dissection and sieving. Fraction 2 (40–100 μm) is usually used in microfluidic devices such as the 3D cell culture chip from AIM Biotech shown, which contains a center gel region. Gel loading port and media ports labeled (B) , along with center and side channels (C) . (I) The AIM Biotech microfluidic device was used by Wang et al. to test a selective RIP1 inhibitor on pancreatic ductal adenocarcinoma PDMEs. After RIP1i treatment ( n = 5 patients shown), PDME live/dead ratio, and size decreased. (A–I) reproduced with permission from Astolfi et al. , Aref et al. , Mazzocchi et al. , and Wang et al. , respectively. (D) Top: **Result for PDMEs stained and imaged a second time; Bottom: * p -value = 0.014. (G) * p < 0.05, ** p < 0.01. (I) * p < 0.05.

Article Snippet: The spheroid fraction was mixed with collagen-I and used for culture in a cyclic olefin co-polymer (COC)-based 3D microfluidic device (DAX-1, AIM BIOTECH) (Aref et al., ).

Techniques: Derivative Assay, Injection, Laser Capture Microdissection, Confocal Microscopy, In Situ, Incubation, In Vitro, Dissection, Cell Culture, Labeling, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Addressing Patient Specificity in the Engineering of Tumor Models

doi: 10.3389/fbioe.2019.00217

Figure Lengend Snippet: Tumor-on-chip systems. (A) Diagram illustrating the processing steps involved in the preparation of patient-derived tumor cells and tumor-infiltrating lymphocytes (TILs) from the same tumor sample, and live imaging by confocal microscope. (B) 12-channel multiplexed cyclic-olefin-copolymer (COC)-based microfluidic device with V-trap design for capturing tumor sample in flow stream and dual port entry for TILs. (C) A 3D multicellular tumor microenvironment microfluidic model consisting of a middle hydrogel channel (2) surrounded by two media channels (1, 3) for the mechanistic study of the effect of monocytes on T cell receptor-redirected T cell (TCR T cell) killing of tumor cell aggregates. Human monocytes were inserted together with target HepG2-preS1-GFP cell organoids in collagen gel in the central hydrogel region (2), while hepatitis B virus (HBV)-specific TCR T cells were added into one fluidic channel (1) to mimic the intrahepatic carcinoma environment. (D) Representative confocal image of a target cell organoid (in green) surrounded by monocytes (in blue) and HBV-specific TCR T cells (in white), in which the presence of dead target cells is DRAQ7+. (E) Left: Representative target cell HepG2 cell organoids (Hep) cultured with monocytes (Mo) and/or HBV-specific T cell (Ts), in which the presence of dead target cells is DRAQ7+(in red). HBV-specific TCR T cells are labeled with Cell tracker violet dye (in white), while monocytes are unlabeled. Right: Box plot of the percentage of dead target volume after 24 h of co-culture with retrovirally transduced (Tdx) HBV-specific TCR T cells (Tc = control T cell). (F) Metastasis-on-a-chip device and in situ tumor and tissue construct biofabrication. Arrows show fluid flow (E, endothelium; Lu, Lung; C, colorectal cancer organoids made of RFP tagged HCT116 cells; Li, liver; blank, control). Constructs comprised of cells in ECM hydrogels exist under fluid flow and have the capability to experience circulating cells either interact or pass by. (G) Metastasis tracking at day 1 and day 15 showing HCT116 cells colonizing other organs, using phase and epifluorescence microscopy (A–G) reproduced with permission from Lee et al. , Moore et al. , and Aleman and Skardal , respectively. * P ≤ 0.05, *** P ≤ 0.001.

Article Snippet: The spheroid fraction was mixed with collagen-I and used for culture in a cyclic olefin co-polymer (COC)-based 3D microfluidic device (DAX-1, AIM BIOTECH) (Aref et al., ).

Techniques: Derivative Assay, Imaging, Microscopy, Cell Culture, Labeling, Co-Culture Assay, In Situ, Construct, Epifluorescence Microscopy

Journal: Micromachines

Article Title: Microfluidic Mixing and Analog On-Chip Concentration Control Using Fluidic Dielectrophoresis

doi: 10.3390/mi7110214

Figure Lengend Snippet: ( a ) Microfludic device with embedded electrodes. The solute and solute-free streams are delivered to the top and bottom inlets respectively. The electrokinetic mixer, boxed in blue, is shown in a magnified brightfield image; ( b ) Montage of the dual-stage microfluidic device. The upstream stage consists of an electrokinetic mixer which is fed continuously downstream to a gradient generator. An electric field is used to inject two solute streams into a central solute-free stream. This stream then serves as an input to the downstream gradient generator. Scale bar, 100 μm.

Article Snippet: The polymer microfluidic device, which sits atop the glass coverslip was fabricated in PDMS (Momentive, Freemont, CA, USA, RTV 615A).

Techniques:

Journal: Micromachines

Article Title: Microfluidic Mixing and Analog On-Chip Concentration Control Using Fluidic Dielectrophoresis

doi: 10.3390/mi7110214

Figure Lengend Snippet: ( a ) Left: The microfluidic device consists of two stages: an upstream mixing stage with embedded electrodes and a downstream gradient chamber. The microchannel network is designed such that when no alternating current (AC) field is applied, only the low conductive central red stream flows to the gradient chamber. Right: A magnified micrograph of the mixing section. A yellow line depicts the spatial location where a 3D cross-sectional image was capture, shown under the zoomed in image; ( b ) When an AC electric field was applied, mixing occurred and influence the solute concentration supplied to the gradient chamber. The mixing section shows that while the red stream enters the mixing zone and exits with a slight orange hue due to mixing with the two outer streams. Scale bar, 200 μm.

Article Snippet: The polymer microfluidic device, which sits atop the glass coverslip was fabricated in PDMS (Momentive, Freemont, CA, USA, RTV 615A).

Techniques: Concentration Assay

Journal: Micromachines

Article Title: Microfluidic Mixing and Analog On-Chip Concentration Control Using Fluidic Dielectrophoresis

doi: 10.3390/mi7110214

Figure Lengend Snippet: A confocal montage captures fluid motion down the axial length of the mixing channel captured at different positions along the channel height. ( a ) Bottom surface of the microfluidic device. Upon entering the array with an applied voltage of 20 MHz at 1 MHz, the outer green streams inject into the center occupying the entire bottom of the channel; ( b ) 1/3 from the bottom of the device; ( c ) 2/3 from the bottom of the device; ( d ) Top of the microchannel device. Scale bar, 50 μm.

Article Snippet: The polymer microfluidic device, which sits atop the glass coverslip was fabricated in PDMS (Momentive, Freemont, CA, USA, RTV 615A).

Techniques:

Journal: Micromachines

Article Title: Microfluidic Mixing and Analog On-Chip Concentration Control Using Fluidic Dielectrophoresis

doi: 10.3390/mi7110214

Figure Lengend Snippet: Magnified view of the microfluidic gradient chamber. Images were captured at varying time points after the electric field was applied over a period of 5 s. When a 5 MHz, 20 V pp AC electric field is applied upstream it modifies the concentration of mixed solution that enters the gradient chamber. ( a ) When the electric field is off, the connecting gradient side channel lacks any fluid from the tri-laminar outer streams. However, when the field is switched on a chemical gradient forms across the connecting channel, which increases with over a time period of 5 s; ( b ) 1 s; ( c ) 3 s and ( d ) 5 s. Scale bar, 40 μm.

Article Snippet: The polymer microfluidic device, which sits atop the glass coverslip was fabricated in PDMS (Momentive, Freemont, CA, USA, RTV 615A).

Techniques: Concentration Assay