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Poly L lysine coating, protocol, information, for cell culture

Neuvitro Corporation owns the leading technology to provide professional coating with the broadest coverage of substrates, such as PDL, PLL, PLO, Collagen, Gelatin, Laminin, Fibronectin, PDL+Laminin, PDL + Fibronectin, PLO+Laminin, PLO+Fibronectin, PDL+Laminin+Fibronectin, etc. The majority of coated German coverslips available from Electron Microscopy Science, ThermoFisher Scientific, and VWR is actually manufactured by Neuvitro Corporation. Free samples are available.

Primary neuron culture on Neuvitro coating of Poly L Lysine on German coverslips

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

 

Options of pre-coated German coverslips with other coating substrates

 

Below is a List of poly-L-lysine (PLL) needed for coating dishes, plates, and glass coverslips

Cell culture devices, Total coating area per plate, Poly-L-lysine needed

6 well culture plate57cm²427.5ug poly-L-lysine at 50ug/ml

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

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

48 well culture plate45.6cm²342ug poly-L-lysine at 50ug/ml

96 well culture plate30.7cm²230ug poly-L-lysine at 50ug/ml

35mm dish 9cm²  67.5ug poly-L-lysine at 50ug/ml

60mm dish 21cm² 157.5ug poly L lysine at 50ug/ml

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

150mm dish 152cm² 1140ug poly-L-lysine at 75ug/ml

245mm dish 500cm² 3750ug poly-L-lysine at 100ug/ml

12 mm coverslips round 2.3cm² 17.25ug poly-L-lysine at 100ug/ml

15 mm coverslips round 3.5cm² 26.25ug poly L lysine at 100ug/ml

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

22 mm coverslips round 7.6cm² 57ug poly L lysine at 100ug/ml

25 mm coverslips round 9.8cm² 73.50ug poly-L-lysine at 100ug/ml

 

Questions and Answers:

 

Should I dissolve poly-L-lysine in dd-water for coating?

  • Poly  L lysine 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 PLL, the ph value is further reduced to 3, which is not optimized for PLL coating. 

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

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

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

  • Minimum required amount of PLL 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 PLL is 510x7.5ug = 3825ug = 3.825mg.

Collection of poly l lysine coating prtocols:

Poly l lysine coating protocol from Sigma

Optimal conditions must be determined for each cell line and application.

  1. Add 50 mL of sterile tissue culture grade water to 5 mg of poly-lysine.

  2. Aseptically coat culture surface with 1.0 mL/25 cm2 (only). Rock gently to ensure even coating of the culture surface.

  3. After 5 minutes, remove solution by aspiration and thoroughly rinse surface with sterile tissue culture grade water.

  4. Allow to dry at least 2 hours before introducing cells and medium.

Poly l lysine coating protocol from ResearchGate:

  1. wash coverslips with Ethanol (100%)

  2. wash coverslips H2O

  3. let coverslips dry

  4. coat coverslips for 5 min with120ul poly-L-Lysin solution (MW 70,000-120,000: 0.01%, Sigma)

  5. wash 3 times with H2O

  6. let coverslips dry

Poly l lysine coating protocol from Salmon Lab

  1. 1. Coat acid-washed coverslips in bulk with 10-15 mL 1 mg/mL PLL in ddH20 in a petri dish. Rock for at least 30 min at room temperature or overnight in the cold room.

  2. 2. Wash coverslips in ddH20 at least 10x (free polyaminoacids are cytotoxic!)

  3. 3. Rinse coverslips in 100% EtOH and allow to dry by resting on one end in an open tissue culture dish in a sterile incubator or hood. Add cells when dry.

  4. 4. For longer storage, allow coverslips to dry on a piece of filter paper. Store dry in a sterile tissue culture dish for several months. Perform step 3 before use.

Poly l lysine coating protocol from PITT

  1. Place glass coverslips in 6-well dish using special forceps for coverslips

  2. Add 2 ml of poly-L-lysine (0.1% from Sigma, diluted 1:10 in sterile water) to each well. Make sure coverslip is submerged. Incubate 15 min.

  3. Wash 3x with sterile PBS

  4. Place under the hood UV light 10-15 min to sterilize (remove cover). Don’t expose the medium to UV!!!

  5. Plate cells as usual (use 2-2.5 ml for each well).

Poly l lysine coating protocol from WISC

  1. Slide Coating (polylysine treatment)Date During all procedures wear powder free gloves and wash fresh gloves with soap and water.This protocol is for coating 60 slides.1.Load 60 slides (GoldSeal) into stainless steel slide racks (30 slides/rack). Check each slidefor scratches and defects. Discard any questionable slides.

  2. Prepare cleaning solution as follows: 350ml/slide rackFor 350mlFor 700ml50g NaOH100g NaOHLot# 100ml MQ H2O200ml MQ H2ONaOH (g) 250ml 95% EtOH500ml EtOHDissolve NaOH in water and slowly add EtOH (Caution EXOTHERMIC Rx)Add additional MQ H2O until solution is clear (about 100ml/350ml solution)

  3. Add 350ml of cleaning solution to each glass dish with slides. Cover dishes with glass andshake gently for 2 hrs.

  4. Remove slides from cleaning solution and plunge into 400ml fresh MQ water in dish. Plungeup and down vigorously and shake for 5 minutes.

  5. Repeat this wash step 5 times with fresh MQ water for each wash. It is imperative that allNaOH is removed before polylysine coating. X X X X X

  6. Prepare polylysine solutionFor 350mlFor 700ml 35ml polylysine (Sigma)70ml polylysineLot# 35ml 1X PBS (filtered)70ml 1X PBSExpiration Date 280ml MQ H2O560ml MQ H2O

  7. Add 350ml of polylysine solution to a clean glass dish and submerge washed slides (in rack)and shake gently for 1hr.

  8. Remove slides from polylysine and rinse by plunging 10X into fresh MQ H2O.9.Spin slides in slide rack at 500 RPM, 5 minutes, wrap slides in aluminum foil and placeslides in 42oC oven for 2-3 hrs

  9. At this point you want to minimize exposure of slides to dust. Working in a hood, transferslides to clean slide box and label box with the date that the slides were coated and name ofperson performing coating.**Slides must be aged at least 2 weeks prior to printing. The aging allows slides to becomesufficiently hydrophobic so that printed DNA is maintained in compact spot.

List of commercially available PLL 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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

  • 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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