Plasma cleaned glass coverslips (22 × 22 mm No. 1) were aminosilanized through Vectabond treatment (Vectabond in acetone 2% vol/vol; Vector Laboratories). Silicone templates (Grace bio-Labs) were used to treat a small section of the coverslips with a PEG solution containing 5kDa biotin-terminated PEG (2.5% w/w in molecular biology grade (MB) water, NOF Corp.), and 5kDa mPEG succinimidyl carbonate (25% w/w in MB water, Laysan Bio Inc.) in 0.1M sodium bicarbonate (Sigma-Aldrich). The coverslips were then left to incubate in a dark and moist environment overnight. On the day of the experiment, the coverslips were treated with another round of PEGylation with a short chain 333 Da NHS-ester PEG (Thermo Scientific) and incubated for 2–3 hrs. After washing off excess PEG, the coverslips were dried with a mild flow of nitrogen. Custom hybriwell chambers (Grace bio-Labs) with dual silicon press-fit tubing connectors (Grace bio-Labs) were placed atop the coverslips to construct a flow chamber.
Silicone templates
Manufactured by Grace Bio-Labs
Silicone templates are versatile tools used in various laboratory applications. They provide a stable and customizable surface for various experiments and procedures. These templates are made of high-quality silicone, offering durability and heat resistance. They can be used to create specific shapes or patterns as required by the research or production needs.
Automatically generated - may contain errors
Lab products found in correlation
Vectabond treatment, by Vector Laboratories (3 mentions)
Dual silicon press fit tubing connectors, by Grace Bio-Labs (3 mentions)
Sodium bicarbonate, by Merck Group (3 mentions)
Vectabond, by Vector Laboratories (2 mentions)
Nhs ester peg, by Thermo Fisher Scientific (2 mentions)
Custom hybriwell chambers, by Grace Bio-Labs (2 mentions)
5kda mpeg succinimidyl carbonate, by Laysan Bio (2 mentions)
3 protocols using silicone templates
1
PEGylation of Glass Coverslips for Flow Chambers
2
Silanization and PEGylation of Coverslips
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Coverslips (22 × 22 mm No. 1) were washed with Liqui-Nox phosphate-free detergent (Alconox Inc.) and 4.3% NH4OH and 4.3% H2O2. Coverslips were then plasma cleaned using a Harrick Plasma PDC-32G Plasma Cleaner and then aminosilanized through Vectabond treatment (Vector Laboratories). A circular area on the slide was then isolated using Silicone templates (Grace bio-Labs) and treated with a PEG solution containing 5 kDa biotin-terminated PEG (2.5% w/w in molecular biology grade (MB) water, NOF Corp.), and 5 kDa mPEG succinimidyl carbonate (25% w/w in MB water, Laysan Bio Inc.) in 0.1 M sodium bicarbonate (Sigma-Aldrich) overnight in a dark and moist environment. On the day of the experiment, the coverslips were washed with PBS, treated with short chain 333 Da NHS-ester PEG (Thermo Scientific) and incubated for 2–3 h. Slides were then washed and dried with N2 gas. A chamber was then constructed over the treated circular area by applying hybriwell chambers (Grace bio-Labs) then dual silicon press-fit tubing connectors (Grace bio-Labs).
3
PEGylation of Glass Coverslips for Flow Chambers
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