Micropatterns to control individual cell shape and adhesion pattern were produced as previously described (Azioune et al., 2009 (link)). Briefly, glass coverslips (22 × 22 mm No. 1.5, VWR) were activated with plasma (Zepto Plasma System, Diener Electronic) for 1 min and incubated with 0.1 mg/ml PLL(20)-g[3,5]-PEG(2) (SuSoS) in 10 mM HEPES at pH 7.4, for 1 h, at RT. After rinsing and air-drying, the coverslips were placed on a synthetic quartz photomask (Delta Mask), previously activated with deep-UV light (PSD-UV, Novascan Technologies) for 5 min; 3 µl of MiliQ water were used to seal each coverslip to the mask. The coverslips were then irradiated through the photomask with the UV lamp for 5 min. Afterward, coverslips were incubated with 25 μg/ml FBN (Sigma-Aldrich) and 5 μg/ml Alexa 546– or 647–conjugated fibrinogen (Thermo Fisher Scientific) in 100 mM NaHCO3 at pH 8.6, for 1 h, at RT. Cells were seeded at a density of 50,000 cells/coverslip and allowed to spread for ∼10–15 h before imaging. Nonattached cells were removed by changing the medium ∼2–5 h after seeding.
Alexa 546 or 647 conjugated fibrinogen
Manufactured by Thermo Fisher Scientific
Alexa 546– or 647–conjugated fibrinogen is a fluorescently labeled protein product used in various research applications. The core function of this product is to provide a labeled form of the glycoprotein fibrinogen, which can be used to study its behavior and interactions in experimental settings.
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Lab products found in correlation
2 protocols using alexa 546 or 647 conjugated fibrinogen
1
Micropatterning for Cell Shape Control
2
Micropatterning Cells with Fibronectin
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Micro-patterns to control individual cell shape and adhesion pattern were produced as previously (Azioune et al. 2009). Briefly, glass coverslips (22 X 22mm No. 1.5, VWR) were activated with plasma (Zepto Plasma System, Diener Electronic) for 1 min and incubated with 0.1 mg/ml of PLL(20)-g[3,5]-PEG(2) (SuSoS) in 10 mM HEPES at pH 7.4, for 1 h, at RT. After rinsing and air-drying, the coverslips were placed on a synthetic quartz photomask (Delta Mask), previously activated with deep-UV light (PSD-UV, Novascan Technologies) for 5 min. 3 µl of MiliQ water were used to seal each coverslip to the mask. The coverslips were then irradiated through the photomask with the UV lamp for 5 min. Afterwards, coverslips were incubated with 25 μg/ml of fibronectin (Sigma-Aldrich) and 5 μg/ml of Alexa546 or 647-conjugated fibrinogen (Thermo Fisher Scientific) in 100 mM NaHCO3 at pH 8.6, for 1 h, at RT. Cells were seeded at a density of 50.000 cells/coverslip and allowed to spread for ~10-15h before imaging. Non-attached cells were removed by changing the medium ~2h-5h after seeding.
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