Artel's 10 Tips To Improve Your Pipetting Technique

Artel
22 Dec 200908:51
EducationalLearning
32 Likes 10 Comments

TLDRThis script emphasizes the importance of proper pipetting techniques to ensure accurate and reliable laboratory results. It outlines ten key tips, including pre-wetting pipette tips to reduce sample loss, working at a uniform temperature for volume consistency, examining tips for droplets, using standard mode for most transfers, pausing after aspiration for liquid stabilization, pulling pipettes straight out to avoid sample variance, minimizing pipette handling to maintain temperature equilibrium, immersing tips to the correct depth, selecting the right pipette tips for a tight seal, and maintaining consistent plunger speed and pressure for reproducible results. These practices are crucial for building confidence in asset results and ensuring liquid handling quality in the lab.

Takeaways
  • 💧 Pre-wet the pipette tip at least three times to increase humidity and reduce sample loss due to evaporation.
  • 🌡️ Allow liquids and the pipette to equilibrate to ambient temperature to minimize volume variation.
  • 🧐 Examine the tip before dispensing and wipe any droplets with a lint-free cloth to avoid sample loss.
  • 🔄 Use standard mode pipetting for most sample transfers for better accuracy and precision.
  • ⏱️ Pause consistently for one to two seconds after aspiration to let the liquid stabilize inside the tip.
  • 🚫 Pull the pipette straight out of the container without touching the sides to maintain volume consistency.
  • 👐 Minimize handling of the pipette and tips to prevent body heat from disrupting temperature equilibrium.
  • 📏 Immerse the tip to the proper depth, staying clear of the container walls and bottom, to ensure accurate sample volume.
  • 🔩 Use the correct pipette tip for the pipette to ensure a tight seal and minimize sample leakage.
  • 🛠️ Maintain consistent plunger speed and pressure during each sample transfer for reproducible results.
Q & A

  • What is the primary purpose of pre-wetting a pipette tip?

    -Pre-wetting a pipette tip increases the humidity within the tip air space, which helps reduce sample loss due to evaporation before delivery.

  • How many times should you pre-wet the pipette tip to ensure proper wetting?

    -You should pre-wet the pipette tip at least three times to increase humidity in the tip and reduce variations in volume delivery.

  • Why is it important to work at temperature equilibrium when pipetting?

    -The volume of sample delivered can vary with environmental conditions such as room temperature, so allowing liquids and the pipette to equilibrate to ambient temperature minimizes variation and ensures accurate dispense volume.

  • What should you do if you notice liquid droplets on the outside of the pipette tip after aspirating a liquid?

    -If there are liquid droplets on the outside of the tip, you should carefully dab the droplets with a lint-free cloth, avoiding wiping near the tip opening to prevent sample loss due to wicking.

  • What is the recommended pipetting mode for most sample transfers?

    -For most sample transfers, it is recommended to use the standard mode pipetting technique, which provides better accuracy and precision when transferring aqueous solutions.

  • Why is it advised to pause consistently after aspiration when pipetting?

    -A brief pause after aspiration allows the liquid and the pressure inside the tip to stabilize, minimizing errors that can result from the liquid bouncing slightly when the plunger stops.

  • How should you pull the pipette out of the container after aspirating a sample?

    -You should pull the pipette straight out of the container without touching the sides, ensuring that the tip is held vertically to avoid wicking or loss of sample.

  • What is the impact of minimizing handling of the pipette and the tip?

    -Minimizing handling reduces heat transfer to the pipette, which can disrupt temperature equilibrium and cause sample volume errors. It also helps maintain the integrity of the pipette and tips.

  • How deep should the pipette tip be immersed when aspirating a sample?

    -The pipette tip should be immersed two to six millimeters below the meniscus, avoiding contact with the container walls and bottom to prevent excess droplets and ensure accurate sample volume.

  • Why is it crucial to use the correct pipette tip for the pipette being used?

    -Using the correct pipette tip ensures a tight seal between the pipette and the tip, which minimizes sample loss due to leakage and provides more dependable sample delivery.

  • What is the recommended technique for depressing and releasing the plunger during pipetting?

    -The plunger should be depressed and released smoothly, with consistent pressure and speed, to avoid inconsistencies in the technique that could allow air to creep into the assay or process.

Outlines
00:00
🧪 Proper Pipetting Techniques for Accurate Sample Handling

This paragraph discusses the importance of using proper pipetting techniques to ensure accurate and consistent sample handling in a laboratory setting. It emphasizes the need for pre-wetting pipette tips to increase humidity and reduce sample loss due to evaporation. It also highlights the significance of working at temperature equilibrium to minimize volume variations and the importance of examining the pipette tip for any droplets before dispensing the sample. The paragraph provides practical tips such as using standard mode pipetting for most transfers, pausing consistently after aspiration to allow liquid stabilization, and pulling the pipette straight out of the container without touching the sides to avoid sample loss. These techniques are crucial for building confidence in asset results and ensuring liquid handling quality in a lab.

05:02
🌡️ Minimizing Errors in Pipetting Through Temperature Control and Proper Handling

The second paragraph focuses on minimizing pipetting errors by controlling the temperature and handling of the pipette and its tip. It advises against overhandling the pipette to prevent body heat from disrupting temperature equilibrium, which could lead to sample delivery issues. The paragraph also stresses the importance of immersing the pipette tip to the correct depth during aspiration to avoid excess droplets and ensure accurate sample volume. Additionally, it highlights the need to use the correct pipette tip for the specific pipette to ensure a tight seal and dependable sample delivery. The use of consistent plunger speed and pressure is also emphasized for reproducible results. By following these guidelines, one can build confidence in their work and trust the results obtained through pipetting.

Mindmap
Keywords
💡Pipe Pinning Technique
Pipe pinning technique refers to the process of pre-wetting a pipette tip before pipetting to increase the humidity within the tip's air space. This technique is crucial for reducing sample loss due to evaporation. In the video, it is emphasized that pre-wetting the tip at least three times helps to ensure volume delivery consistency and build confidence in the results.
💡Humidity
Humidity in the context of the video script refers to the amount of moisture in the air space of a pipette tip. Maintaining an appropriate level of humidity is important because it helps to prevent sample evaporation, which can lead to inaccurate volume delivery. The script suggests that pre-wetting the pipette tip increases humidity, thus reducing sample loss.
💡Equilibration
Equilibration in the script denotes the process of allowing liquids and the pipette to reach the same temperature as the lab environment before use. This is essential because the volume of a liquid can change with temperature, affecting the accuracy of pipetting. The video emphasizes the importance of working at a uniform temperature to minimize variations in dispensed volume.
💡Environmental Conditions
Environmental conditions, such as room temperature, significantly impact the volume of sample delivered by a pipette. The script highlights that these conditions can cause variations in liquid volume, thus it is recommended to allow liquids and pipettes to equilibrate to ambient temperature to ensure accurate pipetting.
💡Liquid Handling Quality
Liquid handling quality pertains to the precision and accuracy with which liquids are transferred using pipettes. The video script underscores the importance of using proper pipetting techniques to ensure high liquid handling quality, which in turn helps to trust the assay results and maintain consistency in laboratory work.
💡Standard Mode Pipetting
Standard mode pipetting is a technique where the plunger is depressed to the first stop, the tip is immersed into the sample, and the plunger is released to aspirate. This method is recommended for most sample transfers in the script. It is suggested that using standard mode for pipetting aqueous solutions helps achieve better accuracy and precision.
💡Reverse Mode Pipetting
Reverse mode pipetting is an alternative technique mentioned in the script where the plunger is depressed past the first stop, and then released after immersing the tip into the sample. This results in aspirating more volume than desired, which can be useful for viscous or volatile solutions. The script indicates that reverse mode may be the preferred method in certain contexts.
💡Consistent Plunger Speed and Pressure
Consistent plunger speed and pressure are vital for reproducible results when using pipettes. The script advises that having a steady and smooth movement of the plunger during both aspiration and delivery ensures accuracy and minimizes the risk of air entering the assay or process.
💡Immersion Depth
Immersion depth refers to how deeply the pipette tip is inserted into the liquid during aspiration. The script instructs to immerse the tip two to six millimeters below the meniscus, avoiding contact with container walls and bottom, to prevent sample contamination and ensure accurate volume transfer.
💡Pipette Tip
A pipette tip is a disposable plastic component that attaches to the end of a pipette to hold liquid samples. The script emphasizes the importance of using the correct pipette tip for the pipette being used, as mismatching or using poor-quality tips can result in inadequate seals and sample leakage.
💡Sample Loss
Sample loss in the context of the script refers to the reduction in the amount of sample due to various factors such as evaporation, wicking, or improper pipetting techniques. The video provides several tips to minimize sample loss, such as pre-wetting the pipette tip, working at temperature equilibrium, and using proper pipetting techniques.
Highlights

Pre-wetting the pipette tip before pipetting increases humidity and reduces sample loss due to evaporation.

Proper pre-wetting technique involves aspirating and dispensing liquid at least three times to wet the tip.

Working at temperature equilibrium minimizes variations in volume delivery.

Allowing liquids and pipettes to equilibrate to ambient temperature is crucial for accurate pipetting.

Examining the pipette tip before dispensing can prevent sample loss due to droplets on the exterior.

Using a lint-free cloth to dab excess droplets from the pipette tip avoids sample contamination.

Standard mode pipetting is recommended for most sample transfers for better accuracy and precision.

Reverse mode pipetting is suitable for viscous or volatile solutions and can help prevent sample loss.

Consistent plunger speed and pressure after aspiration help stabilize the liquid inside the pipette tip.

Pausing one to two seconds after aspiration allows for liquid stabilization, especially with viscous solutions.

Pulling the pipette straight out of the container prevents sample loss due to surface tension effects.

Minimizing handling of the pipette and tips reduces heat transfer and maintains temperature equilibrium.

Immersing the pipette tip to the proper depth prevents excess liquid from clinging to the exterior.

Using the correct pipette tip ensures a tight seal and minimizes sample loss due to leakage.

Smooth and consistent plunger movement during sample transfer is key to reproducible results.

Proper pipetting techniques build confidence in work and trust in results, ensuring liquid handling quality.

Transcripts

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Related Tags
Lab TechniquesLiquid HandlingPipette TipsSample AccuracyPre-wettingTemperature EquilibriumVolume DispensePipetting AccuracyLab EquipmentQuality AssuranceStandard Pipetting