The principle of polylysine coating cell attachment is that the polycationic polylysine molecules adsorb strongly to glass surfaces, leaving cationic sites which combine with the anionic sites on cell surfaces. Polylysine is a cationic polymer. Under physiological conditions, the amine group on polylysine molecules are protonated. Its cationic nature makes it an ideal coating substrate for the adhesion of negatively charged biomolecules and adhesion of cells. The cell surface is known to be negatively charged. Thus, polylysine coatings are widely used for the adhesion of cells in cell culture.

 

List of available Polylysine coated German coverslips for cell culture (1/2 year storage life)

 

Options of pre-coated German coverslips with other coating substrates

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Questions and Answers:

 

Should I dissolve polylysine in dd-water for coating?

• polylysine coating in dd-water is simple but the coating efficiency is low. The pH of dd-water is acidic, usually lower than pH6. After dissolving polylysine, the ph value is further reduced to 3, which is not optimized for polylysine coating. 

• Solution: Use high pH polylysine coating, such as pH 8.5, in borate buffer, which enhances polylysine adsorption to cell culture surface. 

• Critical: The borate buffer residue must be rinsed away thoroughly from coverslips before cell culture.

How much poly lysine needed to coat my dish / plates / coverslips?

• Minimum required amount of polylysine is 7.5ug/cm² surface area of your culture dish or coverslips. For example, to coat 100 pieces of 18mm diameter coverslips, the total surface area of 100 pieces of coverslips is 0.9x0.9x3.1416 x2x100 = 510cm². The required polylysine is 510x7.5ug = 3825ug = 3.825mg.

 

Polylysine coating protocol

Polylysine solutions are typically used to coat glassware and plasticware  for the attachment and growth of cells.

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Preparation

1. Prepare a polylysine stock solution by dissolving 100 mg polylysine in 100 ml water and filter sterilize through a 0.22-micron filter.

2. Store polylysine in 5-ml aliquots at -20°C.

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Polylysine coating culture dishes, multiwell plates, or chamber slides

1. When ready to use, dilute 1 part polylysine stock solution with 19 parts water to prepare a 50 g/ml working polylysine solution.

2. Fill tissue culture dishes, multiwell plates, or slide wells with the polylysine working solution and incubate 1 hr in a 37°C incubator, then remove polylysine solution by vacuum aspiration and allow surface to dry.

3. To coat coverslips: Sterilize coverslips by autoclaving or by incubating them in 95% ethanol and drying before polylysine coating. Place coverslips in a single layer in a petri dish containing polylysine working solution and incubate 1 hr at 37°C. Remove coverslips using sterile forceps and allow surface to dry.

4. Store polylysine coated tissue culture ware up to 3 months at 4°C. Use diluted polylysine solutions only once.

5. When coating with polylysine, the efficiency of coating is relatively low when prepared with distilled water due to the acidic pH of the solution that can range between a pH of 3.3-6.6. One recommendation [1] to improve coating is to use an 8.5  pH borate buffer to dissolve polylysine, with thorough rinsing after coating. In addition to the basic techniques shown here, there are a variety of approaches recommended by vendors.

 

There are two kinds of polylysine available as coating substrate to anchor cells in culture. Poly-L-lysine and poly-D-lysine. You should choose poly-D-lysine instead of poly-L-lysine to coat your plates, slids, or coverslips because some cells in culture can actually digest poly-L-lysine gradually.

 

Below is a List of polylysine needed for coating dishes, plates, and glass coverslips

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Cell culture devices, Total coating area per plate, polylysine needed

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6 well culture plate57cm²427.5ug polylysine at 50ug/ml

12 well culture plate45.6cm²342ug polylysine at 50ug/ml

24 well culture plate45.6cm²342ug poly lysine at 50ug/ml

48 well culture plate45.6cm²342ug polylysine at 50ug/ml

96 well culture plate30.7cm²230ug polylysine at 50ug/ml

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35mm dish 9cm²  67.5ug polylysine at 50ug/ml

60mm dish 21cm² 157.5ug polylysine at 50ug/ml

100mm dish 55cm² 412.5ug poly lysine at 75ug/ml

150mm dish 152cm² 1140ug polylysine at 75ug/ml

245mm dish 500cm² 3750ug polylysine at 100ug/ml

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12 mm coverslips round 2.3cm² 17.25ug polylysine at 100ug/ml

15 mm coverslips round 3.5cm² 26.25ug polylysine at 100ug/ml

18 mm coverslips round 5cm² 37.50ug poly lysine at 100ug/ml

22 mm coverslips round 7.6cm² 57ug polylysine at 100ug/ml

25 mm coverslips round 9.8cm² 73.50ug polylysine at 100ug/ml

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• List of commercially available polylysine coated coverslips:

• H-12-pll:  12 mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L-lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $54.75

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• GG-12-pll: 12 mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $89.50

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• H-12--1.5-pll: 12 mm diameter, # 1.5 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $59.75

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• GG-12-1.5-pll: 12 mm diameter, # 1.5 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $99.50

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• H-14-pll: 14 mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $54.75

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• GG-14-pll: 14 mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $89.50

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• H-15-pll: 15 mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $54.75

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• GG-15-pll: 15 mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $89.50

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• H-18-pll: 18 mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $56.25

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• GG-18-pll: 18mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $92.50

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• H-18-1.5-pll: 18mm diameter, # 1.5 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $61.25

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• GG-18-1.5-pll: 18mm diameter, # 1.5 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $102.50

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• H-20-pll: 20mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $56.25

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• GG-20-pll: 20mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $92.50

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• H-22-pll: 22mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $56.25

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• GG-22-pll: 22mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $92.50

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• H-22-1.5-pll: 22mm diameter, # 1.5 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $61.25

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• GG-22-1.5-pll: 22mm diameter, # 1.5 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $102.50

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• H-24-pll: 24mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $57.75

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• GG-24-pll: 24mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $95.50

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• H-25-pll: 25mm diameter, # 1 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $59.75

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• GG-25-pll: 25mm diameter, # 1 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $99.50

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• H-25-1.5-pll: 25mm diameter, # 1.5 thickness, 45 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $72.75

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• GG-25-1.5-pll: 25mm diameter, # 1.5 thickness, 90 pieces of CE certified crystal clear German glass coverslips with Poly-L- lysine (PLL) coating on both sides for cell culture and confocal fluorescence microscopy, sealed 15 pieces per bag, sterilized, ready to use $125.50

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• PDL coating protocol

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