Improving the efficiency of coating plates and flasks with collagen

  • Improving the efficiency of coating plates and flasks with collagen

    Coat your Petri dishes with ease

    Coating dishes or tissue culture flasks with extracellular matrix proteins, such as collagen or fibronectin, is a common procedure in cell culture laboratories. However, coating large numbers of dishes is a tedious, time-consuming task that is repetitive for the user. This application note describes how using the DOSE IT results in significant time savings and reduces excess strain compared to a regular pipette controller.

  • Table of contents

    Coating dishes or tissue culture flasks with extracellular matrix proteins, such as collagen or fibronectin, is a common procedure in cell culture laboratories. However, coating large numbers of dishes is a tedious, time-consuming task that is repetitive for the user. This application note describes how using the DOSE IT results in significant time savings and reduces excess strain compared to a regular pipette controller.

Key benefits

  • The DOSE IT allows fast and accurate coating of culture flasks or dishes without excessive strain on the user
  • CUSTOMIZE mode enables easy and efficient creation of personalized liquid handling protocols
  • The DOSE IT has a very small footprint and weighs only 3.5 kg, making it perfect for benchtop or in-hood dispensing applications
  • A large display and clear user guidance support fast set-up and use
  • The DOSE IT is an intuitive, programmable peristaltic pump that simplifies the dispensing of cell culture media, buffers and other solutions for volumes ranging from 0.1 ml to 2500 ml

Overview: How to coat Petri dishes with collagen

All three protocol steps involving liquid transfer were analyzed by measuring the time taken to process 100 Petri dishes, using either a pipette controller in combination with 10 ml serological pipettes or the DOSE IT peristaltic pump.

Before coating the Petri dishes, a collagen stock solution should be prepared according to the manufacturer’s instructions. This stock solution should be diluted to a final concentration of 50 μg/ml collagen in 0.02 M acetic acid.

Overview of the steps:

  1. DOSE IT set-up
  2. Coating the Petri dishes with collagen
  3. Removing the excess collagen solution
  4. Washing the coated Petri dishes

Step-by-step procedure:

1. DOSE IT set-up

Tubing adjustment, priming and calibration for dispensing the solution

Open the pump head, adjust the tubing clamps and insert sterilized 4.0 mm tubing into the DOSE IT. Set the flow rate to 470 ml/min (280.7 rpm) for optimal performance. Put the aspiration tube with the tube collar into the collagen solution, and the dispensing tube into an empty bottle to fill the empty tubing with liquid.

Perform the priming and calibration steps according to the Operating instructions.

2. Coating the Petri dishes with collagen

Dispensing the collagen solution into the Petri dish

Put the dispensing tube above the Petri dish. The DOSE IT directly dispenses the required volume of collagen solution into the Petri dish (Table 1). Make sure the entire bottom of the Petri dish is covered with collagen solution. After filling all the Petri dishes, remove the aspiration tube from the collagen solution and put the dispensing tube into an empty bottle. Incubate the Petri dishes at room temperature for one hour. The DOSE IT can also be used for other culture flasks or dishes, with dispensing volumes given in Table 1.

Tip:

  • Optimizing collagen concentration and coating times for your cell lines and experimental needs is recommended.

Table 1: Typical coating and washing volumes.

Labware Recommended coating volume (ml) Recommended washing volume (ml)
6 well 0.6 1.0
35 mm PD 2.0 1.0
60 mm PD 5.0 2.5
90 mm PD 10.0 5.0
T25 8.0 4.0
T75 24.0 12.0

 

3. Removing the excess collagen solution

Aspirating the collagen solution from the Petri dish

After incubation, carefully aspirate the solution from each of the Petri dishes into an empty bottle (Figure 1). The excess collagen solution can be collected in a bottle and reused up to a maximum of 5 times, if desired. Remove the aspiration tube from the used collagen solution bottle, put the dispensing tube into an empty bottle to drain, then drain the collagen solution from the tubing.

Coating and Washing Petri dishes using DOSE IT peristaltic pump
Figure 1: Aspirating collagen solution.

4. Washing the coated Petri dishes

Rinsing the Petri dishes with PBS buffer to remove the remaining acid

Put the aspiration tube into the PBS buffer (or serum-free medium), and the dispensing tube into an empty bottle to refill the empty tubing. Fill the tubing with PBS buffer, then rinse the Petri dish twice with PBS to remove the remaining acid (see Table 1 for volumes). Air dry the Petri dishes at room temperature inside a laminar flow hood, then seal with parafilm and store at 4 °C until needed.

Results

For all steps, using the DOSE IT was significantly faster than working with a serological pipette controller, with an overall time saving of 40 % (Table 2).

Table 2: Measured time for handling 100 Petri dishes using a serological pipette controller or a DOSE IT peristaltic pump. Righthand column shows % time saving using a peristaltic pump.

  Pipette controller DOSE IT Time saving (%)
Dispense collagen 13 min 6 min 54
Aspirate collagen 15 min 12 min 20
Washing (2x) 60 min 35 min 40
Total 88 min 53 min 40

 

Conclusion

  • For all steps, using the DOSE IT was significantly faster than working with a serological pipette controller. When working with a serological pipette controller, all the liquid transfers require a two-step procedure. In contrast, when working with the DOSE IT the same liquid transfer becomes a one-step process.
  • In addition to the time savings, the DOSE IT also provides ergonomic benefits. The pipette connector attached to the silicone tubing allows the user to attach any size of aspirator or Pasteur pipette, as required for the individual application.
  • The DOSE IT’s connector is compact and lightweight, weighing less than 50 grams. This makes it possible to work more ergonomically, and closer to the labware, saving the user from continuously lifting the long serological pipette connected to a much heavier pipette controller. Since the need for repetitive hand movements is eliminated, the risk of repetitive strain injury is minimized.

Any questions? I'm happy to help!

  • Éva Mészáros, PhD

    Application Specialist

    Email

Instruments and accessories

DOSE IT Laboratory Peristaltic Pump Benchtop switch Foot switch Silicone tubing Aspiration/dispensing tubes Tube collar

DOSE IT Laboratory Peristaltic Pump

DOSE IT has been especially designed for laboratories looking for an easy-to-use, compact and portable peristaltic pump. With its light weight and compact design, DOSE IT fits everywhere in the lab and can be easily moved. The intuitive user interface with straightforward instructions makes it very easy to operate. DOSE IT is especially helpful for dispensing culture media, buffers or other solutions.

Part No. 171 000

Find out more about DOSE IT Laboratory Peristaltic Pump
DOSE IT peristaltic pump

Benchtop switch

DOSE IT Benchtop switch: For ease of operation, with connecting cable.

Part No. 171 081

Find out more about accessories for DOSE IT
Benchtop switch for ease of operation, with connecting cable

Foot switch

DOSE IT Foot switch: For hands-free operation, with connecting cable.

Part No. 143 200

Find out more about accessories for DOSE IT
Foot switch for hands-free operation, with connecting cable

Silicone tubing

DOSE IT silicone tubing: 2.5 m, autoclavable tubing, ID (MM) 1 and 4.

Part No. 171 021
Part No. 171 024

Find out more about accessories for DOSE IT
Silicone tubing, lengt 2.5m, 6mm inner diameter

Aspiration/dispensing tubes

DOSE IT aspiration/dispensing tube, length 10 cm, stainless steel, one end dented, ID (MM) 2 and 4.

Part No. 171 052
Part No. 171 054

Find out more about accessories for DOSE IT
Aspiration/dispensing tube, length 35 cm, stainless steel, one end dented, 6mm inner diameter

Tube collar

Tube collar used as a weight for the aspiration tube or as a clamp spacer for the dispensing tube. ID (MM) 1-3.

Part No. 171 071

Find out more about accessories for DOSE IT
Tube collars used as a weight for the aspiration tube

Customer’s voice

I Love my DOSE IT. The DOSE IT is an absolute must have for a laboratory in need of efficient, repeat dispensing protocols. I use it to rapidly add precise volumes of media for enriching samples and it saves me so much time!

JOE HIRSCH, Smithfield Foods Corp Lab