Journal: Communications Biology

Article Title: The cancer glycocalyx mediates intravascular adhesion and extravasation during metastatic dissemination

doi: 10.1038/s42003-021-01774-2

Figure Lengend Snippet: A Schematic diagram of the microfluidic device used to perfuse MVNs under a constant pressure difference Δ p and cells being carried by luminal flow into A narrow channels and B impacting the endothelium at bifurcations or C large vessels (partially realized with Biorender.com). B Speed of inert beads carried by luminal flow driven by different pressure differences ( n = 50 beads tracked in three devices, average and standard deviation shown) and arrest efficiency of TCs ( n = 4 devices, average and standard deviation shown) as a function of the pressure difference. C Confocal image of two arrest mechanisms showing MDA-MB-231 cells within MVNs arrested by (1) physical trapping and (2) adhesion. The scale bar is 200 µm. D Bead speed through the MVNs ( n as above, the error bars indicate the standard deviation) and vessel diameter as a function of specific GCX component removal (the error bars indicate the standard deviation between the averages of n = 3 devices, 3 regions of interest each). E Arrest efficiency of TCs as a function of GCX enzymatic treatment of the MVNs alone, TCs alone, or both MVNs and TCs ( n = 4 devices). F Cumulative percentage of TCs either physically trapped in small vessels or adhered to large vessels in the MVNs ( n > 40 cells). Statistical significance was assessed by student’s t test assuming normally distributed data, p < 0.05 *, p < 0.01 **, p < 0.0001 ****. A normal distribution of the data in ( B ) and ( D ) was confirmed by Kolmogorov–Smirnov test.

Article Snippet: Extravasation experiments were conducted in MVNs cultured within AIM microfluidic chips (AIM Biotech, gel channel width of 1.3 mm and 250 μm height).

Techniques: Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: Phthalimide Derivative Shows Anti-angiogenic Activity in a 3D Microfluidic Model and No Teratogenicity in Zebrafish Embryos

doi: 10.3389/fphar.2019.00349

Figure Lengend Snippet: 3D angiogenesis assay in microfluidic device. (A) Representative confocal images of the angiogenic sprouting in the 3D extracellular matrix-like hydrogel. Human endothelial cells were treated with Thalidomide or one of the compounds at 0.5, 25, and 100 μM for 72 h. Treatment with vehicle was used as control. Cells are stained for F-actin and Hoechst to visualize actin cytoskeleton filaments and nuclei, respectively. (B) Analysis of the volumetric sprouting for each condition. The box and whiskers plots show all the data points quantified for at least three devices per condition. Statistics were calculated by one-way ANOVA. (C) The different network complexity for each compound or for Thalidomide was analyzed by quantifying the branch levels and the number of segments for each branch level combining all the concentrations for each compound. n.s. = not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Microfluidic plastic chips and holders were purchased from AIM Biotech company (AIM Biotech, Singapore).

Techniques: Angiogenesis Assay, Staining

Journal: Neuron

Article Title: Multimodal Single-Cell Analysis Reveals Physiological Maturation in the Developing Human Neocortex

doi: 10.1016/j.neuron.2019.01.027

Figure Lengend Snippet: (A) Schematic of experimental workflow using the Polaris microfluidic system.

Article Snippet: The same physiological profiles that we observed in culture were recapitulated on microfluidic chips , allowing us to use the combined mRNA and response profiling as a “Rosetta Stone” to translate between physiological profiling and gene expression datasets.

Techniques:

Journal: Neuron

Article Title: Multimodal Single-Cell Analysis Reveals Physiological Maturation in the Developing Human Neocortex

doi: 10.1016/j.neuron.2019.01.027

Figure Lengend Snippet: (A) Single-cell Ca2+ responses to different agonists analyzed on microfluidic chips and on physiological rig were co-clustered to reveal physiological types (P).

Article Snippet: The same physiological profiles that we observed in culture were recapitulated on microfluidic chips , allowing us to use the combined mRNA and response profiling as a “Rosetta Stone” to translate between physiological profiling and gene expression datasets.

Techniques:

Journal: Advanced Science

Article Title: Pericytes Promote More Vascularization than Stromal Cells via an Interleukin‐6‐Dependent Mechanism in Microfluidic Chips

doi: 10.1002/advs.202408131

Figure Lengend Snippet: Replication of the two main vascularization processes in microfluidic devices. A) Main mechanisms of vessel formation in the human body. B) A schematic drawing of the microfluidic chip shows the internal hydrogel channel (blue) and the surrounding parallel media channels (red). C) Fluorescence images showing freshly seeded endothelial cells (Vybrant DiD, magenta) and supporting cells (Vybrant DiO, yellow), which can be either pericytes or stromal cells, in two different set‐ups to study vasculogenesis (left) and angiogenesis (right). White arrows mark the interface between the hydrogel and media channel where the endothelial cells attach. Scale bars: 500 µm.

Article Snippet: To compare the processes of vasculogenesis and angiogenesis ( Figure 1 A ), and how pericytes and stromal cells act in both, we used a commercially available three‐channel microfluidic chip from AIM Biotech, Singapore (Figure ).

Techniques: Fluorescence

Journal: Advanced Science

Article Title: Pericytes Promote More Vascularization than Stromal Cells via an Interleukin‐6‐Dependent Mechanism in Microfluidic Chips

doi: 10.1002/advs.202408131

Figure Lengend Snippet: Comparison of the pro‐angiogenic potential of pericytes and stromal cells. A) Immunofluorescence images of the angiogenic sprouting within the microfluidic chips co‐cultured with either pericytes or stromal cells, stained for nuclei (DAPI; cyan), CD31 showing the endothelial vessel‐like structures (magenta) and the supporting cells’ F‐actin cytoskeleton (Phalloidin; yellow). F‐actin is not visible within the endothelial cells since it was subtracted from the picture during the image processing to increase clarity. Scale bar: 200 µm (left), 100 µm (right). B–E) Comparison of total vessel volume ( p = 0.0428), length ( p = 0.0340), branching points ( p = 0.0329), and average diameter ( p = 0.6508) between both co‐culture conditions (n = 3, unpaired t‐test, p < 0.05 is considered significant). F) Comparison of the distance between supporting cells and the nearest vessel‐like structure ( p < 0.0001). The trunked violin plots depict summary statistics and the kernel density estimation to show the frequency distribution of each condition. The middle line represents the median (n = 3, Mann‐Whitney test, p < 0.05 is considered significant). G) Comparison of supporting cells’ average sphericity ( p = 0.2667, n = 3, unpaired t‐test, p < 0.05 is considered significant). Pericytes and stromal cells from 3 different donors each were used for the co‐cultures (n = 3), while the endothelial cell donor remained constant. Asterisks indicate a statistical significance (* p < 0.05, **** p < 0.0001).

Article Snippet: To compare the processes of vasculogenesis and angiogenesis ( Figure 1 A ), and how pericytes and stromal cells act in both, we used a commercially available three‐channel microfluidic chip from AIM Biotech, Singapore (Figure ).

Techniques: Comparison, Immunofluorescence, Cell Culture, Staining, Co-Culture Assay, MANN-WHITNEY

Journal: Advanced Science

Article Title: Pericytes Promote More Vascularization than Stromal Cells via an Interleukin‐6‐Dependent Mechanism in Microfluidic Chips

doi: 10.1002/advs.202408131

Figure Lengend Snippet: Comparison of pericytes and stromal cells in promoting vasculogenesis of endothelial cells. A) Immunofluorescence images of the vasculogenesis process within the microfluidic chips co‐cultured with either pericytes or stromal cells, stained for nuclei (DAPI; cyan), CD31 showing the endothelial vessel‐like structures (magenta) and the supporting cells’ actin cytoskeleton (F‐actin, Phalloidin; yellow). Scale bars: 200 µm. B) Comparison of total vessel volume between both co‐culture conditions ( p = 0.0223, n = 3, unpaired t‐test, p < 0.05 is considered significant). C) Comparison of the distance between each type of supporting cell and the nearest vessel‐like structure. The trunked violin plots depict summary statistics and the kernel density estimation to show the frequency distribution of each condition. The middle line represents the median (n = 3, Mann‐Whitney test, p < 0.05 is considered significant). D) Comparison of the final total number of endothelial cells (ECs; p = 0.1372, n = 3, unpaired t‐test, p < 0.05 is considered significant) and supporting cells (SPs; p = 0.999, n = 3, Mann‐Whitney test, p < 0.05 is considered significant). E) Comparison of supporting cells’ average sphericity ( p = 0.2099, n = 3, unpaired t‐test, p < 0.05 is considered significant). F) Quantification and comparison of ten cytokines tightly related to vascularization, namely angiopoietin 1, angiopoietin 2, vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor 2 (FGF‐2), interleukin‐6 (IL‐6) and interleukin‐8 (IL‐8), tumor necrosis factor (TNF), PECAM‐1 and placenta growth factor (PlGF). Statistical values can be found in Table (Supporting Information) (n = 3, p < 0.05 is considered significant). Pericytes and stromal cells from 3 different donors each were used for the co‐cultures (n = 3), while the endothelial cell donor remained constant. Asterisks indicate statistical significance (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: To compare the processes of vasculogenesis and angiogenesis ( Figure 1 A ), and how pericytes and stromal cells act in both, we used a commercially available three‐channel microfluidic chip from AIM Biotech, Singapore (Figure ).

Techniques: Comparison, Immunofluorescence, Cell Culture, Staining, Co-Culture Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Cytosolic delivery of large supramolecular protein complexes arranged on DNA nanopegboards

doi: 10.1101/236729

Figure Lengend Snippet: (a) Principle of microfluidic cell squeezing. A cell suspension containing the DONs (black rectangles) is squeezed through a microchannel constriction smaller than the cell diameter. Transient formation of pores in the cell membrane enables diffusion of the DONs into the cytosol. Schematic illustrations and representative AFM images of (b) DON-1 containing 6 Cy3/Cy5 FRET pairs; (c) DON-2 containing 12 biotin (Btn) and 6 Cy5 modifications for binding of up to 12 streptavidin (STV) proteins; (d) DON-3 containing 6 chlorohexyl (CH) and 6 Cy5 modifications for binding of up 6 HOB-tagged FLIP proteins; (d) DON-4 containing 6 CH, 4 Btn and 6 Cy5 modifications for binding of up to six molecules of HOB-tagged FLIP and four STV protein molecules. The percentage of occupied protein binding sites is given underneath the AFM images. Scale bars are 100 nm. ( f ) Schematic drawing of the glucose biosensor FLIP-HOB. Binding of glucose leads to the decrease in FRET and a concomitant change in the fluorescence intensity ratio I(527 nm)/I(476 nm). The HOB domain is used for ligation with chlorohexyl (CH)-modified DNA molecules.

Article Snippet: For the micromechanical transfection of functionalized DON by squeezing, 1.10 5 HeLa cells were suspended in 100 μL of a 10 nM DON solution in TEMg or PBS and were transferred to the squeezing device (SQZbiotech) combined with microfluidic chips (SQZbiotech) with channel dimensions of 10 × 7, 10 × 8 or 10 × 9 (length / μm × diameter / μm).

Techniques: Diffusion-based Assay, Binding Assay, Protein Binding, Fluorescence, Ligation, Modification