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Sulfo sanpah solution

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Sulfo-SANPAH solution is a heterobifunctional crosslinking reagent that can be used to immobilize proteins and other biomolecules onto surfaces. It contains an N-hydroxysulfosuccinimide (sulfo-NHS) ester group that can react with primary amine groups, and a photoreactive nitrophenyl azide group that can form covalent bonds with surfaces upon UV irradiation.

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Lab products found in correlation

Fibronectin, by Thermo Fisher Scientific (2 mentions) Collagen, by Merck Group (2 mentions) F8806, by Thermo Fisher Scientific (1 mentions) Rheowin data manager software, by Thermo Fisher Scientific (1 mentions) Haake mars 3, by Thermo Fisher Scientific (1 mentions) Sylgard 184, by Dow (1 mentions)

4 protocols using sulfo sanpah solution

1

Fabrication and Characterization of PDMS Substrates

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PDMS substrates of varying rigidity was prepared based on the previous literature.[12] The SYLGARD components A and B (Sylgard 184; Dow Corning, Midland, MI, USA) were mixed at 50:1 and 100:1 ratios and spread on cell culture vessels (10‐ or 15‐cm‐diameter dish, 48‐ or 6‐well plate) or coverslips (13 mm), and cured at 70 °C overnight. Sulfo‐SANPAH solution (0.1 mg mL−1, 22 589; Thermo Scientific, Waltham, MA, USA) was dropped on the surface of the PDMS substrate, irradiated for 10 min with UV light, and irradiated again for 5 min after the removal of the Sulfo‐SANPAH solution. The PDMS substrate was then coated with collagen I (25 µg mL−1) after washing twice with phosphate‐buffered saline (PBS). The mechanical properties of the PDMS substrates were characterized using a rotational rheometer (HAAKE MARS III; Thermo Scientific). Parallel plates were used in oscillation mode (0.1–10 Hz, 25 °C) and the results were analyzed using RheoWin Data Manager software (Thermo Scientific).
Liu Z., Wang L., Xu H., Du Q., Li L., Wang L., Zhang E.S., Chen G, & Wang Y. (2020). Heterogeneous Responses to Mechanical Force of Prostate Cancer Cells Inducing Different Metastasis Patterns. Advanced Science, 7(15), 1903583.
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2

Protocol for PAA Gel Preparation and Functionalization

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The protocol of PAA gel preparation followed a previous report (Wang and Pelham, 1998 (link)) with some modifications. In brief, the surface of a glass-bottom culture dish was processed by NaOH (0.1 M, 5 min), (3-Aminopropyl) trimethoxysilane (100%, 3 min), and glutaraldehyde (0.5%, 20 min) to facilitate the covalent attachment of PAA. The PAA gel was prepared by mixing ddH2O (258 µl), acrylamide (40%, 17 µl), bis-acrylamide (2%, 12 µl), ammonium persulfate (10%, 1.5 µl), and N,N,N’,N’-Tetramethylethylenediamine (100%, 0.45 µl), and 15 µl of the mixture was then mixed with 1.4 µl fluorescent microspheres (F8809, F8806; Invitrogen), dropped onto a previously prepared coverglass, and flattened with another cover glass. The dish was then put onto the surface of 24°C water under lamp light for polymerization. 20 min later, the coverglass over the gel was removed and the gel was exposed to UV for activating PAA after adding sulfo-SANPAH solution (1 mM; 22589; Thermo Fisher Scientific). The solution was removed 10 min later, and the gel was washed with Hepes (50 mM) on a shaker for 30 min, three times. The gel-coated dish was used immediately after preparation.
Jiang J., Zhang Z.H., Yuan X.B, & Poo M.M. (2015). Spatiotemporal dynamics of traction forces show three contraction centers in migratory neurons. The Journal of Cell Biology, 209(5), 759-774.
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3

Fabrication of Microfluidic Organ-on-Chip Device

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The device was fabricated using polydimethylsiloxane (PDMS; SYLGARD® 184 silicone elastomer kit) with previously described soft lithography techniques21 (link),64 (link) or obtained from Emulate, Inc. The chip design, which was similar to previously published devices21 (link), included apical and basal microchannels with dimensions of 1 × 1 × 16.7 mm and 1  × 0.2 × 16.7 mm (w x h x l), respectively, that were separated by a 50 μm thick porous PDMS membrane (7μm diameter holes with 40μm spacing). Both channels were washed with 70% ethanol, filled with 0.5 mg/mL sulfo-SANPAH solution (Thermo Fisher Scientific, A35395) in 50mM HEPES pH 8 and placed under a UV lamp (Nailstar, NS-01-US) for 20 minutes to activate the surface.The microchannels were then rinsed sequentially with 50 mM HEPES buffer and cold PBS, and then filled with a coating solution of PBS containing 100 μg/ml fibronectin (Gibco, 33016-015) and 50 μg/ml collagen (Sigma, C2249) and placed at 37°C for 2 hours. The solution was then aspirated from the chip and it was allowed to air dry overnight.
Chou D.B., Frismantas V., Milton Y., David R., Pop-Damkov P., Ferguson D., MacDonald A., Bölükbaşı Ö.V., Joyce C.E., Teixeira L.S., Rech A., Jiang A., Calamari E., Jalili-Firoozinezhad S., Furlong B.A., O’Sullivan L.R., Ng C.F., Choe Y., Clauson S., Myers K.C., Weinberg O.K., Hasserjian R.P., Novak R., Levy O., Prantil-Baun R., Novina C.D., Shimamura A., Ewart L, & Ingber D.E. (2020). On-chip recapitulation of clinical bone-marrow toxicities and patient-specific pathophysiology. Nature biomedical engineering, 4(4), 394-406.
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4

Fabrication of Microfluidic Organ-on-Chip Device

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The device was fabricated using polydimethylsiloxane (PDMS; SYLGARD® 184 silicone elastomer kit) with previously described soft lithography techniques21 (link),64 (link) or obtained from Emulate, Inc. The chip design, which was similar to previously published devices21 (link), included apical and basal microchannels with dimensions of 1 × 1 × 16.7 mm and 1  × 0.2 × 16.7 mm (w x h x l), respectively, that were separated by a 50 μm thick porous PDMS membrane (7μm diameter holes with 40μm spacing). Both channels were washed with 70% ethanol, filled with 0.5 mg/mL sulfo-SANPAH solution (Thermo Fisher Scientific, A35395) in 50mM HEPES pH 8 and placed under a UV lamp (Nailstar, NS-01-US) for 20 minutes to activate the surface.The microchannels were then rinsed sequentially with 50 mM HEPES buffer and cold PBS, and then filled with a coating solution of PBS containing 100 μg/ml fibronectin (Gibco, 33016-015) and 50 μg/ml collagen (Sigma, C2249) and placed at 37°C for 2 hours. The solution was then aspirated from the chip and it was allowed to air dry overnight.
Chou D.B., Frismantas V., Milton Y., David R., Pop-Damkov P., Ferguson D., MacDonald A., Bölükbaşı Ö.V., Joyce C.E., Teixeira L.S., Rech A., Jiang A., Calamari E., Jalili-Firoozinezhad S., Furlong B.A., O’Sullivan L.R., Ng C.F., Choe Y., Clauson S., Myers K.C., Weinberg O.K., Hasserjian R.P., Novak R., Levy O., Prantil-Baun R., Novina C.D., Shimamura A., Ewart L, & Ingber D.E. (2020). On-chip recapitulation of clinical bone-marrow toxicities and patient-specific pathophysiology. Nature biomedical engineering, 4(4), 394-406.
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