Journal: Developmental cell

Article Title: Long-term high-resolution imaging of developing C. elegans larvae with microfluidics

doi: 10.1016/j.devcel.2016.11.022

Figure Lengend Snippet: Device layout and operating principle. (A) Schematic of a worm chamber with inlet and outlet ports for the flow layer (red) and the inlet port for the pressure layer (blue). Features in light gray are for structural support only. (B) Side view (top) and top view (bottom) of a worm chamber; Scale bar 250 μm; side view not drawn to scale. Height of flow layer is 20 μm. Height of the pressure layer chamber is 50 μm. Left: Top: Between immobilizations, the pressure layer is kept under vacuum such that the height of the worm chamber is increased. Bottom: The animal is confined in the chamber through an array of posts with 4.88 μm spacing. Entrance to the chamber is 10 μm wide. The animal is drawn in cross-sectional view. (C) Illustration of the immobilization procedure. Left: gradually increasing suction at the flow layer outlet (bottom) pushes the animal to the side of the chamber without flushing it through the small channels (top & middle). Right: vacuum in the pressure layer is gradually released and replaced by pressure. This slowly pushes down the membrane between the pressure and flow layers to gently compress and immobilize the worm. (D) Differential Interference Contrast (DIC) image of an immobilized L2 larva in the chamber. Inset: Magnified view of the mid-body region. Arrows show nuclei of two vulval precursor cells as well as the anchor cell (AC). Dashed line indicates gonad outline. (E) Illustration of the microfluidic chip with 10 identical worm chambers. Features in light gray are for structural support. (F) To achieve short imaging intervals, immobilization procedures and imaging for chambers 1,3,…9 and 2,4,…10 are staggered using two independent pressure/vacuum pipelines. See also Figures S1 and S2.

Article Snippet: Mold fabrication We designed the flow and compression layers of the microfluidic device in AutoCAD (Autodesk) ( File S1 ).

Techniques: Membrane, Imaging

Journal: Developmental cell

Article Title: Long-term high-resolution imaging of developing C. elegans larvae with microfluidics

doi: 10.1016/j.devcel.2016.11.022

Figure Lengend Snippet: Tracking vulval precursor cell (VPC) divisions. (A) Cell divisions in C. elegans vulval development at the L3 stage. Dark ellipses indicate nuclei; brighter outlines indicate cell membranes. VPC progeny denoted by Pn.px where n=3,4…8 and x=a (anterior) or p (posterior) for the first two divisions. P6.p (red) executes the 1° fate (three rounds of cell divisions), P5.p & P7.p (green) execute the 2° fate (three rounds of cell divisions except P5.ppp and P7.paa). P3.p, P4.p and P8.p (blue) divide once and fuse to the hypodermis (3° fate). (B) DIC micrographs of 1-cell, 2-cell, 4-cell, and 8-cell stages of vulval development in a repeatedly immobilized larva. First two rounds of cell division for P5-7.p occur in the anterior-posterior orientation, giving rise to Pn.pxx (n=5–7, x=a,p). The terminal cell divisions of the P6.p granddaughters all occur in the transverse (left-right) orientation. The terminal cell divisions of P5.paa & P5.pap as well as P7.ppa and P7.ppp are longitudinal; P5.ppa and P7.pap undergo a final transverse division and P5.ppp and P7.paa do not divide further. P6.p descendants are outside of the focal plane of bottom panel. Scale bar, 20 μm. (C) DIC micrographs of a P4.p cell division imaged at 8 min intervals. Purple outlines highlight P4.p and progeny. Left: P4.p nucleus and nucleolus are visible. Center left: Nuclear envelope breakdown occurred, metaphase plate has formed and no nuclear components can be seen. This is the most prominent feature of cell division in DIC microscopy and what is scored (Sulston and Horvitz, 1977). Center right: Cytokinetic furrow is apparent by the kink on the ventral side of the cell (purple outline). Right: P4.pa/p nuclei are visible. Scale bar, 20 μm. Division time would be scored as 4h24min. See also Movies S2 and S3. (D–F) Cell cycle statistics of VPC divisions. Dark red/green/blue circles indicate division times or cell cycle length of 1°/2°/3° fate cells, respectively, obtained by following divisions in the microfluidic device (n=106 animals). Light red/green/blue circles same, except obtained through traditional lineaging (n=13). Note that manual lineaging was performed at 23°C and cell division times were rescaled to 20 °C (see Experimental Procedures for details). (D) Time point of the first VPC division relative to the P5.p division in each animal. Note that in one animal, the anchor cell was misplaced anteriorly and P4.p-P6.p adopted vulval fates with cell cycle timings indicative of cell fate. (E) As in D but relative to the P8.p division. (F) Length of the second VPC cell cycle. (G) Length of the third VPC cell cycle.

Article Snippet: Mold fabrication We designed the flow and compression layers of the microfluidic device in AutoCAD (Autodesk) ( File S1 ).

Techniques: Microscopy

Journal: Communications Engineering

Article Title: Microfluidic chip for precise trapping of single cells and temporal analysis of signaling dynamics

doi: 10.1038/s44172-022-00019-2

Figure Lengend Snippet: A Two-layer microfluidic chip design to ( B ) deliver time- and dose-modulated input stimuli to single cells and investigate ( C , D ) apoptosis in K562 cell, activated upon treatment with different doses and time of DMSO and ( E , F ) the translocation dynamics of STAT-1 signaling protein in NIH3T3 cells, when stimulated with type 2 interferons.

Article Snippet: The microfluidic chip was designed in Autodesk AutoCAD 2017.

Techniques: Translocation Assay

Journal: Communications Engineering

Article Title: Microfluidic chip for precise trapping of single cells and temporal analysis of signaling dynamics

doi: 10.1038/s44172-022-00019-2

Figure Lengend Snippet: A The complete setup of two-layer microfluidic chip comprises of external pneumatic membrane valves and pressure source, to actuate on-chip pneumatic membrane valves open or close, and a graphical user interface, written in MATLAB, to provide instructions and aid in automation of experimental workflow. The microfluidic chip is placed in a stage-top incubator and the cells, on-chip, are imaged using fluorescence microscope. B Two-layer microfluidic chip design where flow in sixteen channels (blue) is controlled using eight control lines (red). The inlet channels on the chip are controlled by additional control lines (labeled 9–16) and the outlet is controlled by control port 17. Each flow channel is integrated with cell isolation unit, in green, that has pillar-like traps to physically isolate single cells upon contact. C The pillars, fabricated from PDMS, are separated by a small gap of 4 µm and incorporates a V-shaped cup to efficiently hold individual cells in place. D The control lines, in the top layer, upon orthogonally intersecting the flow lines, in the bottom layer creates several thin pneumatic push-down membrane valves on-chip. The pneumatic membrane valves are actuated open or close through an external pressure source that control the flow of reagents in the channels.

Article Snippet: The microfluidic chip was designed in Autodesk AutoCAD 2017.

Techniques: Membrane, Fluorescence, Microscopy, Labeling, Cell Isolation

Journal: Communications Engineering

Article Title: Microfluidic chip for precise trapping of single cells and temporal analysis of signaling dynamics

doi: 10.1038/s44172-022-00019-2

Figure Lengend Snippet: In this design, each of the sixteen microfluidic channels is integrated with pillar-like traps for trapping of single cells on microfluidic chip. A , B Regardless of whether single cells ( A ) or multiple cells ( B ) are injected at the entrance of the microfluidic channel, simulation model in ANSYS Fluent demonstrated that the cells need to be at the center of the channel to physically interact with the pillars and be isolated. C Flow profile in the channel, when a particle of 15 µm is trapped by the pillars. From the flow profile it can be predicted that when other cells enter the channel, they are diverted to the sides of the pillar by virtue of flow profile as there is almost little to no flow through the 4 µm gap between the pillars. D Once a cell or a particle has been physically trapped by the PDMS pillars, they will be subjected to shear stress, ranging between 0.25 Pa and 2 Pa, from the flow in the channels. (The white space in the particle, that can be seen as hole, is the part of the section that attaches with the trap during assembly. Upon hiding the traps, that specific section in the rigid particles also disappears giving the impression of hole. This section is not affected by the flow, according to the simulation software.) The particle dimension in this simulation model is 15 µm and the flow rate is 1 µL/min.

Article Snippet: The microfluidic chip was designed in Autodesk AutoCAD 2017.

Techniques: Injection, Isolation, Shear, Software

Journal: Communications Engineering

Article Title: Microfluidic chip for precise trapping of single cells and temporal analysis of signaling dynamics

doi: 10.1038/s44172-022-00019-2

Figure Lengend Snippet: A This design has demonstrated high efficiency in trapping suspension K562 cells that can be treated with stimuli to monitor cellular response immediately after seeding in the channels. B This design is also compatible with adherent NIH3T3 cells. Fibroblast cells, when flushed in the channels are round and over the time will sink to the bottom of the channel to stretch which is promoted with fibronectin coating. Additionally, these cells also migrate in and out of the trap and within the channel. C A single fibroblast, that took 1 h, to stretch in the channel and after additional 1 h stretched completely for distinct visualization of the cytoplasm and nucleus. The blue-fluorescent signal is from CFP-labeled IRF7 transcription factor that resides in the cytoplasm until activation. D Highly efficient and consistent trapping of single cells in our device with over 90% channels being able to isolate cells, both adherent and suspension, at low pressures ( N : Number of individual events). At higher pressures, of around 10 kPa, the cells squeezed through the middle of the PDMS pillars and were difficult to retain. Error bar represents mean ± SD. E The cell trapping experiment was repeated multiple times on three different microfluidic chips to determine the design’s reusability and reproducibility. For each individual chip, we observed more than 85 events with single-cell trapping and very few events with more than one cell in channel. Error bar represents mean ± SD. F Representative microscopic images of viable K562 cells NIH3T3 cells that were cultured in microfluidic channels. G K562 cells and NIH3T3 cells, cultured on the chip, showed high viability of over about 80% through media exchange. Error bar represents mean ± SD.

Article Snippet: The microfluidic chip was designed in Autodesk AutoCAD 2017.

Techniques: Suspension, Labeling, Activation Assay, Cell Culture

Journal: Communications Engineering

Article Title: Microfluidic chip for precise trapping of single cells and temporal analysis of signaling dynamics

doi: 10.1038/s44172-022-00019-2

Figure Lengend Snippet: A Translocation was measured by plotting the ratio of fluorescence intensities between the nucleus and the cytoplasm. At different concentrations of 1 and 2 µg/mL distinct translocation patterns, with respect to time, were observed when cells were treated with a single 10 min pulse of IFNγ. B At concentrations of 1 and 2 µg/mL distinct translocation patterns, with respect to time, were observed when cells were continuously exposed to IFNγ which was also different from the patterns observed due to short single pulse exposure. C , D Representative images of NIH3T3 cells that showed STAT-1 activity when treated with 10 min pulse (left) and continuous exposure (right) of IFNγ. E The bar graph displays the percentage of cells demonstrating STAT-1 activation in single cells in microfluidic channel, which was much lower compared to percentage of activated cells in bulk. N = 3. Error bar represents mean ± SD. F Representative image of cells in population study that showed higher percentage of active cells in comparison to single-cell study.

Article Snippet: The microfluidic chip was designed in Autodesk AutoCAD 2017.

Techniques: Translocation Assay, Fluorescence, Activity Assay, Activation Assay, Comparison

Journal: Neural Regeneration Research

Article Title: Nerve growth factor-basic fibroblast growth factor poly-lactide co-glycolid sustained-release microspheres and the small gap sleeve bridging technique to repair peripheral nerve injury

doi: 10.4103/1673-5374.344842

Figure Lengend Snippet: The schematic diagram of the drug screening biomimetic microfluidic chip. bFGF: Basic fibroblast growth factor; CCU: cell culture unit; CGG: concentration gradient generator; NGF: nerve growth factor.

Article Snippet: The microfluidic chip virtual model was designed using Autodesk AutoCAD software.

Techniques: Drug discovery, Cell Culture, Concentration Assay

Journal: Neural Regeneration Research

Article Title: Nerve growth factor-basic fibroblast growth factor poly-lactide co-glycolid sustained-release microspheres and the small gap sleeve bridging technique to repair peripheral nerve injury

doi: 10.4103/1673-5374.344842

Figure Lengend Snippet: Virtual model and schematic of a drug screening biomimetic microfluidic chip. The chip comprises an upstream CGG and downstream parallel CCU, and includes two inlet ports, one liquid outlet, one drug CGG and eight cell culture chambers. CCU: Cell culture unit; CGG: concentration gradient generator.

Article Snippet: The microfluidic chip virtual model was designed using Autodesk AutoCAD software.

Techniques: Drug discovery, Cell Culture, Concentration Assay

Journal: Neural Regeneration Research

Article Title: Nerve growth factor-basic fibroblast growth factor poly-lactide co-glycolid sustained-release microspheres and the small gap sleeve bridging technique to repair peripheral nerve injury

doi: 10.4103/1673-5374.344842

Figure Lengend Snippet: Identification of drug concentration gradients of the biomimetic microfluidic chips. (A) Images 1–8 represent eight concentration gradients (more details of concentrations are shown in ). (B) The difference between the theoretical and experimental data of a concentration gradient generator. Data are expressed as mean ± SD. The study was repeated three times.

Article Snippet: The microfluidic chip virtual model was designed using Autodesk AutoCAD software.

Techniques: Concentration Assay

Journal: Neural Regeneration Research

Article Title: Nerve growth factor-basic fibroblast growth factor poly-lactide co-glycolid sustained-release microspheres and the small gap sleeve bridging technique to repair peripheral nerve injury

doi: 10.4103/1673-5374.344842

Figure Lengend Snippet: Screening of the drug concentration in primary Schwann cells. (A) Live and dead cell staining of rat Schwann cells under eight different NGF/bFGF drug concentrations on the microfluidic chip. Green is AO stained cells (live), red is PI stained cells (dead). The cell number and proliferation rate of Schwann cells gradually increased from the 1 st chamber to the 4 th chamber, and gradually decreased from the 4 th chamber to the 8 th chamber, reaching a peak in the 4 th chamber (22.86 ng/mL NGF combined with 4.29 ng/mL bFGF). Scale bars: 25 μm. (B) Cell proliferation rate (cell number after culture with NGF/bFGF/initially implanted cell number × 100) under eight different concentrations. (C) Cell number after culture with NGF/bFGF under eight different drug concentrations. Data are expressed as mean ± SD. The above experiments were independently repeated three times. * P < 0.05, vs . other groups (one-way analysis of variance followed by Bonferroni post hoc test). 1–8: Cell culture chambers. AO: Acridine orange solution; bFGF: basic fibroblast growth factor; NGF: nerve growth factor; PI: propidium iodide.

Article Snippet: The microfluidic chip virtual model was designed using Autodesk AutoCAD software.

Techniques: Concentration Assay, Staining, Cell Culture

Journal: bioRxiv

Article Title: Spatiotemporal NF-κB dynamics encodes the position, amplitude and duration of local immune inputs

doi: 10.1101/2021.11.30.470463

Figure Lengend Snippet: (A) Diagram illustrating a model of immune signaling following a local infectious signal. A macrophage detects a pathogen, and releases TNF to alert nearby cells. Receiving cells interpret this signal and regulate NF-κB dynamics. (B) Simplified NF-κB network used for mathematical modeling. (C) Microfluidic device designed to study the infection scenario represented in (A). Either TNF (blue dot) or a single macrophage was loaded in the source chamber. The macrophage was stimulated with LPS and washed with fresh medium. When the valve is opened, TNF or macrophage secretions diffuse into the receiving chamber and activate fibroblasts. (D) Diffusion of signals in the receiving chamber is tested with fluorescent dye. (E) Kinetics of dye fluorescence are plotted at various distances from the source chamber. (F) NF-κB activation at each location derived from simulation. Cell activation probability was calculated at various distance from source. Each point represents a single cell with the regulatory network in (C). (G) Simulation shows the release of TNF from source chamber elicits wave-like NF-κB propagation in the receiving cell population. (H) Microfluidic experiments exhibited wave propagation of NF-κB activation after release of TNF (100 ng/ml) from a source chamber. Fluorescently tagged NF-κB shuttles into the nucleus upon activation. Yellow arrows indicate examples of activated cells in each microscopy image. Graphs on the right of each image illustrate the fraction of activated cells at different distances. Circles and error bars indicate the mean and standard deviation in ten replicates. (I) NF-κB responses in near (0 – 0.4 mm), mid (0.8 – 1.2 mm), and far (1.8 – 2.2 mm) regions. Red lines show the mean of all cell responses in each region (n > 300), while thin lines represent 20 random responses. Purple bars indicate the delay time between the initiation of TNF diffusion and onset of local NF-κB activation, while blue bars indicate the activation time. (J) Activation probability at various distance calculated for three different doses of TNF (10, 30 and 100 ng/ml). Each dose had four replicates.

Article Snippet: The patterns for our diffusion microfluidic chip were designed with AutoCAD (Autodesk, Inc.) and KLayout.

Techniques: Infection, Diffusion-based Assay, Fluorescence, Activation Assay, Derivative Assay, Microscopy, Standard Deviation

Journal: bioRxiv

Article Title: Spatiotemporal NF-κB dynamics encodes the position, amplitude and duration of local immune inputs

doi: 10.1101/2021.11.30.470463

Figure Lengend Snippet: (A) Downstream expressions from three different cytokine secretion durations (2, 4, and 8 h) are simulated with our 2-D model. Expression at each grid point is calculated using the corresponding NF-κB dynamics as an input to a gene expression function. Blue indicates genes with rapid degradation (early), while red represents genes with slow degradation (late). (B) Diagram illustrates the schematics of spatially-resolved downstream measurement using a custom microfluidic chip. (C) Cells from three different regions are collected 4 h after releasing TNF from source chamber (30 or 100 ng/ml). After sequencing, the pairwise distance between samples are compared in MDS plot for 1,094 significantly upregulated genes. (D) Using RT-qPCR, the kinetics of A20 (early), CASP4, and RANTES (late) genes are measured for different locations in 100 ng/ml sample. (E) Upregulated genes from sequencing data were clustered using correlation method. The functional characteristics of genes in each cluster were analyzed through enrichment of GO terms and TF (transcription factor) motifs. The numbers to the right of the bar indicate the p -value.

Article Snippet: The patterns for our diffusion microfluidic chip were designed with AutoCAD (Autodesk, Inc.) and KLayout.

Techniques: Expressing, Sequencing, Quantitative RT-PCR, Functional Assay

Journal: Scientific Reports

Article Title: On-Chip Fluorescence Switching System for Constructing a Rewritable Random Access Data Storage Device

doi: 10.1038/s41598-017-16535-7

Figure Lengend Snippet: Illustration of the microfluidic chip used for data encoding and rewriting. ( a ) Schematic drawing of the assembled microfluidic device with inlet and outlet connectors. The entire chip consists of 3 layers. The top cover layer provides a seal, and the middle layer made of plastic film containing flow microchannel are placed over an aldehyde-modified glass substrate on which CSs were immobilized. ( b ) Photograph of the film-based microfluidic chip fabricated for the study.

Article Snippet: The film-based microfluidic chip was designed with the assistance of AutoCAD computer software from Autodesk, Inc. (Santa Barbara, CA).

Techniques: Modification

Journal: Scientific Reports

Article Title: On-Chip Fluorescence Switching System for Constructing a Rewritable Random Access Data Storage Device

doi: 10.1038/s41598-017-16535-7

Figure Lengend Snippet: Rewritable DNA data storage system using fluorescence switching on a microfluidic chip. Each letter uses 8 bits per channel to store its information. Fluorescence images of the intentionally made erroneous text KRIBT, and the rewritten text KRIBB are shown after hybridization and strand displacement processes. We set 50% increase in the relative fluorescence intensity on the microfluidic chip as cut-off level for representing the binary 1. Two DSs for #36, #38 spots and one PS for #39 spot were injected through the inlet connector for data rewriting, respectively.

Article Snippet: The film-based microfluidic chip was designed with the assistance of AutoCAD computer software from Autodesk, Inc. (Santa Barbara, CA).

Techniques: Fluorescence, Hybridization, Injection

Journal: Materials Today Bio

Article Title: Advanced liver-on-chip model mimicking hepatic lobule with continuous microvascular network via high-definition laser patterning

doi: 10.1016/j.mtbio.2025.101643

Figure Lengend Snippet: Concept design of liver-on-chip via femtosecond laser patterning. (A) Schematic illustrations of minimum functional unit of the in vivo liver (i.e., hepatic lobule), which is composed of a dense-hepatocyte tissue (brown) and a microvascular network (red). (B) Construction of the hepatic lobule-like structure in a microfluidic chip. (C) Fabrication process of the liver-on-chip via high-definition (HD) laser patterning. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: A master mold of microfluidic chip was designed in AutoCAD (Autodesk, San Francisco, CA, USA) in order to realize the millimeter-scale hepatic lobule in the central channel of the chip.

Techniques: Functional Assay, In Vivo

Journal: Materials Today Bio

Article Title: Advanced liver-on-chip model mimicking hepatic lobule with continuous microvascular network via high-definition laser patterning

doi: 10.1016/j.mtbio.2025.101643

Figure Lengend Snippet: Construction of continuous microvessels in the laser-patterned microchannels. (A) Schematic illustrations of microvessel formation in the laser-patterned microchannels with different diameters (i.e., Ø50 and Ø80 μm) in the cell-containing hydrogel. RFP-HUVECs were seeded to one side channel of the microfluidic chip. (B) Fluorescence images of RFP-HUVECs (red) in the laser-patterned on days 5 and 9. The cross-view images correspond to the dotted lines (i–iii) in the above image. Scale bars: 200 μm. (C) Quantitative analysis of length of continuous microvessel on days 5 and 9. Data represent the mean ± SD (n = 18/group). ∗ p < 0.05 (two-way ANOVA with the post hoc Tukey's honestly significant difference test). (D) Quantitative analysis of width of microvessel on days 5 and 9. Data represent the mean ± SD (n = 18/group). ∗ p < 0.05 (Student's t -test). Blue lines indicate 50 μm and 80 μm in width of microvessel, respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: A master mold of microfluidic chip was designed in AutoCAD (Autodesk, San Francisco, CA, USA) in order to realize the millimeter-scale hepatic lobule in the central channel of the chip.

Techniques: Fluorescence