Receiving Helpdesk

what common error in handling a micropipette can account for under pipetting

by Fermin Mills Published 3 years ago Updated 2 years ago

A few of these common errors are (these also apply for smaller volume pipettes such as a micropipette): Dried out pipette tips. Pipette tips should be made wet before use by aspirating and dispensing a liquid sample at least three times before beginning the pipetting procedure. Incorrect solution temperature.

Working Too Quickly
After aspirating, failure to pause with the pipette tip in the liquid can lead to under-delivery.

Full Answer

What are the most common pipetting errors?

Working too quickly while pipetting is another common pipetting error. Let the liquid settle into the pipette tip for a second or two after aspirating. Now that you know what they are, you can work on proper pipetting techniques to eradicate laboratory error in liquid handling.

What is the most common error with the micropipette?

Note: The most serious and common error is depressing the plunger to the second stop before filling the micropipette. Avoid this error. For dispensing the liquid follow these steps:

What should I do if my micropipette is not dispensing correctly?

Note: The most serious and common error is depressing the plunger to the second stop before filling the micropipette. Avoid this error. For dispensing the liquid follow these steps: Take the volume of liquid in the tip then place the tip against the sidewall of the container or tube in which the liquid needs to be shifted.

What are the disadvantages of using a manual pipettor?

When using a manual pipettor, variation in pipetting speed can result in uneven liquid volumes on aspirate or dispense. Aspirating too quickly can cause inaccuracies with viscous liquids, lead to air bubbles, or liquid contact with the pipette filter, increasing the chance of contamination.

What common error in handling a pipet can account for pipetting too little reagent into a tube under pipetting )?

Pipetting at incorrect angles. Your pipette should be vertical when aspirating. Working too quickly while pipetting is another common pipetting error. Let the liquid settle into the pipette tip for a second or two after aspirating.

What are the common pipetting errors?

Inconsistent Pipetting Depth Immersing the tip too deep in a liquid can result in droplets on the side of the tip or pressure changes that alter volume. Conversely, using too shallow a depth while aspirating can lead to air in the tip.

What types of errors would be introduced by using a single pipette for all operations in this procedure?

What types of errors would be introduced by using a single pipette for all operations in this procedures? Systematic error.

What are the factors affecting pipette performance?

The factor that has the greatest effect is the temperature difference between the delivery device and the liquid. The air gap (dead air volume) between the liquid surface and the piston experiences thermal expansion effects unique to the case.

How can pipetting errors be prevented?

Practice a smooth and slow motion.Pre-wet the pipette tip.Hold the pipette vertically when drawing in liquid and at a 45-degree angle when dispensing.Immerse the tip slightly into the liquid during aspiration.Touch off the pipette on the sidewall of the container.

What are three important precautions in micropipette use?

Take the following precautions when using a micropipette: run back into the piston. ➢ NEVER let the plunger snap back after withdrawing or expelling fluid; this could damage the piston. ➢ NEVER immerse the barrel of the micropipettor in fluid. ➢ NEVER flame the tip of the micropipettor.

What is the largest source of pipetting problems?

Human error is the largest source of pipetting problems, followed by liquids sticking to the tips, and loss of accuracy when working with viscous liquids (multi-option select question, chart depicts the percent of survey respondents who experienced these various pipetting errors).

What are the common errors done in measuring liquids?

Top 5 Errors in PipettingNot Accounting for the Viscosity of a Sample. ... Dispensing Liquids Too Quickly. ... Contamination By “Double Dipping” ... Cleaning Irregularly—or Not at All. ... Taking a Break Only at the End of a Project. ... How Can You Avoid These Errors?

What is the error of a volumetric pipette?

This page provides a table of typical uncertainty values for the calibrated volume of such glassware....Uncertainties for Volumetric Glassware.ItemVolume (mL)Uncertainty (mL)Transfer pipette “To Deliver”2.000±0.0061.000±0.006Mohr (graduated) pipette10.00±0.05Volumetric flask1000.0±0.3014 more rows•Jun 12, 2008

How do you improve pipetting errors?

5 Tips to Improve Your Pipetting AccuracyPre-rinse your new pipette tips before aspirating a sample for delivery. ... Ensure the tips immersion depth is correct. ... Ensure the micropipette is upright during pipetting. ... Apply consistent plunger pressure and speed. ... Good micropipette.

What causes over pipetting?

Allowing the tip of the pipet to rise above the liquid in the container usually causes the liquid to be sucked into the pipet bulb. A broken or chipped pipet can reduce the amount of liquid held after transfer. This causes too much liquid to be delivered.

Why are pipettes inaccurate?

You Use an Air Displacement Pipette, Most Likely Temperature and pressure affect the volume of the air cushion, which affects pipetting accuracy. Also, volatile solvents can evaporate into the air cushion, which leads to an inaccurate and lower dispensed volume than that displayed on the pipette.

Why is it important to remove pipette at an angle?

In addition, removing the pipette at an angle can cause volume variation due to surface tension effects , especially when pipetting small volumes. Pulling the pipette straight out of the container can minimize error.

What is pipetting technique training?

With pipetting technique training programs, pre- and post-training skills assessment are necessary to gauge improvement in pipetting accuracy and precision.

Why is sample volume altered when the pipette is not temperature equilibrated?

This is because air displacement pipettes are affected by air pressure, relative humidity and vapor pressure of the liquid, all of which are temperature-dependent.

Do laboratorians pipette?

Laboratorians pipette on a daily basis. Yet many have never received formal training on pipetting. And like most repetitive, common tasks, pipetting is often taken for granted. It is important to understand the steps that laboratorians can take to improve pipetting skills.

Is it necessary to train pipette operators?

It is not enough to only train pipette operators. Training is important for laboratorians at all levels, whether they are on-the-bench scientists and technicians or laboratory managers. Process-control-focused training can be beneficial for quality and laboratory managers and supervisors.

Why are pipettes important?

Pipettes are not just fancy handlebars for your tips, they are essential for precisely measuring and dispensing liquids. These standard ‘tools of your trade’ enable you to accurately repeat experiments, validate results, make important comparisons between projects and eventually publish that outstanding paper.

What to do when liquid is aspirated into pipette barrel?

That’s right, disassemble for a deep cleaning to prevent your pipette from becoming the source of contamination. But what about pipettes that pass through many hands without incident? Heavy use will cause parts to wear out more quickly, resulting in a significant decrease in how accurately your liquid samples are measured. Repair may be as simple as occasionally replacing an o-ring, but eliminating contamination is something that takes a proactive approach.

Can heavy use cause o-rings to wear out?

Heavy use will cause parts to wear out more quickly, resulting in a significant decrease in how accurately your liquid samples are measured. Repair may be as simple as occasionally replacing an o-ring, but eliminating contamination is something that takes a proactive approach.

Changing Pipetting Angle

When using a standard micropipette to aspirate liquid, it is important to hold the pipettor upright. Pipetting at an angle past 20º impacts the pressure inside the pipette tip, getting less and less accurate as the angle gets steeper.

Always Upright

The angle of the microPro never changes. Plate to plate, well to well, the microPro stays perfectly vertical no matter who is using it.

Inconsistent Pipetting Depth

One of the most common sources of liquid volume inconsistencies is variation in depth during aspiration. Immersing the tip too deep in a liquid can result in droplets on the side of the tip or pressure changes that alter volume. Conversely, using too shallow a depth while aspirating can lead to air in the tip.

Pipetting Depth Recall

Each step of your protocol can be calibrated for a precise pipetting depth. Tips are always at the same depth while aspirating and dispensing liquids, even across 96 wells.

Poor Tip Seal

A pipette tip that is not properly set on its pipette leads to a poor seal that can dramatically impact volumes. This can become especially problematic on handheld multichannel pipettes when it is more difficult to feel if a tip is correctly seated.

Tip Cartridge System

microPro tips are factory installed to the correct position and pressure every time. Seating tips on the instrument is easy and reliable, even with a distracted user.

Variation in Pipetting Speed

When using a manual pipettor, variation in pipetting speed can result in uneven liquid volumes on aspirate or dispense. Aspirating too quickly can cause inaccuracies with viscous liquids, lead to air bubbles, or liquid contact with the pipette filter, increasing the chance of contamination.

How to check pipetting technique?

If you want to check your pipetting technique, measure the effect that your pipetting has on the process that depends on the quantity of fluid you are pipetting. Repeat the pipetting operation on each of several samples. If there is a broad distribution of results, then your technique needs improvement.

Why are TC pipets calibrated?

These pipets are calibrated to deliver the correct volume of liquid without having to blow it away. In a TC pipet, the pipets are calibrated to contain the correct volume. In this case, the last drop needs to be blown away. Many serological pipets are TC pipets and the last drop should be blown away.

What are the two most common mistakes in a vacuum?

The two most common mistakes are probably setting the tip too deep which creates a vacuum and releasing too quickly. Both mistakes will cause the liquid to rise too quickly and enter the shaft where it could potentially cause corrosion. I find it helpful to carefully watch where the liquid goes.

Why are medication errors so common?

Errors in medication are one of the most common causes of harm in patients. They are a failure in the treatment process that has the potential to harm or results in harm in patients.

What are the most common errors in medication?

Errors in medication are one of the most common causes of harm in patients. They are a failure in the treatment process that has the potential to harm or results in harm in patients. Medication errors include: 1 Delay or omission of administration 2 Unauthorized administration of medication 3 Incorrect dosage 4 Administration via the incorrect route, incorrect rate, or with an incompatible fluid or a conflicting medication 5 Medication calculation error 6 Administering to the wrong patient; or 7 Allergy related factors

Can you make mistakes in Quora?

Oh yes, you can make mistakes when using those as well. Edit : List item 1 has been corrected based on some valuable education from Quora User. 1. The first thing is to be aware whether the last drop in your pipet is to be blown away or not.

Can you go too deep in a gel?

Go too deep and tip can puncture the gel. Go too fast and the sample can leak out. Go too shallow and the sample won't enter the well and diffuse away. It's usually helpful to practice loading empty samples with loading dye to get a gauge of what depth to go and how slowly you need to depress the tip.

What is a micropipette?

A Micropipette is a standard type of laboratory equipment that is used to measure the small volumes of liquids. Micropipettes are also used to transfer a small volume of liquids accurately from one vessel to another.

What is the difference between a pipette and a micropipette?

Pipettes and micropipettes are used to measure and deliver an accurate volume of liquid. Pipettes are used to measure small volumes starting from 1 millimeter whereas micropipettes are used to measure the very small volume of liquid usually in microliters.

What is a blue micropipette tip?

Blue micropipette tips are used for P 1000 micropipettes. These tips are used to take a high liquid volume from 200 µ to 1000 µ. These are the micropipettes tips that can take in liquid volume even in milliliters.

How does a micropipette work?

Micropipette works by displacing the air from the pipette shaft and allow the liquid to be drawn into the resulting vacuum. Pipettes and micropipettes are very essential tools used in microbiology labs, medical labs, and environmental sciences for various academics and research studies.

Why are micropipettes used?

Micropipettes are used to measure and transfer the volume of liquid with great accuracy and precision in a laboratory. Accuracy in micropipette measurement is necessary as any discrepancy in volume dispensed can affect the result of the experiment.

What is a pipette barrel?

Pipette barrel is a tube filled with air present with the pipette tip. Pushing the plunger forces out a volume of air and releasing the plunger lets that air come back in the barrel. There are two stops in the plunger which are used differentially for taking up liquid in forward and reverse pipetting.

What is an air displacement micropipette?

Air displacement micropipettes are precise and very accurate micropipettes used to measure a liquid volume range from 0.1 – 5000 µ. These micropipettes are available in different sizes from P 2 to P 5000 as mentioned above. These pipettes are used to measure and transfer the volume of liquid with the help of disposable micropipette tips. Micropipettes tips are available in different colors and sizes and are compatible with specific micropipettes.

How to pre-wet a pipette?

To pre-wet your tip, draw up the liquid into your pipette and then dispense it back into the original vessel. The coating is on the tip. Now, when you draw up the liquid again and dispense it into the receiving vessel, none of it will be left on the tip and lost to wetting.

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

Pre-wetting the tip before you pipette will help increase your accuracy.

Why are positive displacement pipettes so expensive?

The lack of air cushion also decreases the chance of contamination when pipetting corrosives and bio-hazardous material , which makes positive displacement pipettes more suitable for working with those reagents. These pipettes are expensive because the barrel and the tip are a unit and both are replaced when pipetting.

How does an air displacement pipette work?

You Use an Air Displacement Pipette, Most Likely. An air displacement pipette works a bit like a syringe, except that there is an air-filled cushion between the piston and the sample. The air cushion prevents the piston from coming into contact with the solution.

Why is air cushion important in pipetting?

Keeping the solution and the pipette barrel apart is good, but it also puts some limitations on the pipette. Temperature and pressure affect the volume of the air cushion, which affects pipetting accuracy.

Can you use an air displacement pipette?

If you’re working with corrosives or bio-hazardous material, this can be a problem. 2. Don’t Use an Air Displacement Pipette (Depending on Your Application!)

Can liquids roll down into pipette barrels?

There’s nothing preventing the liquid from rolling down into the pipette barrel (remember only air separates the liquid from the pipette barrel). Any liquid inside the pipette barrel results in contamination at the least and can cause some serious corrosion damage.

What are the differences in pipetting technique?

Differences in technique can be attributed to a variety of factors, but it’s important to recognize what’s at stake. Incorrect, incomplete or carelessly performed pipetting steps can – and do – directly impact test results, and might even affect patient outcomes.

How does manual pipetting benefit?

Like most skills combining science with art, manual pipetting really does benefit from regular practice and attention to the finer points. Keep the plunger speed and pressure consistent.

What is the best way to pipette aqueous solutions?

Forward, or standard, pipetting mode is generally the best method to use with aqueous solutions. Reverse-mode pipetting can be useful for viscous or volatile liquids, but it generally over-delivers when using solutions similar to water.

How to aspirate a sample?

Go with the flow 1 Keep the tip in a vertical position during aspiration, and as it exits the source container. 2 Avoid contact between the tip and the container wall during or after aspiration. 3 When aspirating, immerse tips to the right depth – too shallow, and you’ll risk introducing air into the sample. But go too deep and you’ll increase the chance of extra liquid adhering to the outside of the tip. 4 As the sample is dispensed into the receiving container, the tip should not be immersed at all. This will help avoid both over-delivery and cross-contamination.

How to avoid compromising important test results?

To avoid compromising important test results, make sure you’re using the right tools for the job. Use only high-quality pipette tips. Examine the tip before dispensing. Use the right pipette size. Any of these factors could potentially introduce error into test results, and multiple variations only compound the risk.

How to minimize errors in a chemistry lab?

Errors are easily minimized by paying attention to environmental influences, and taking the necessary steps to monitor and control them. Maintain temperature equilibrium while you work. Allow time for liquids, instruments and ambient air temperatures to stabilize. Release your grip.

Should you wipe your tip after aspirating?

Pause briefly – and consistently – after aspirating, while tip remains immersed. If at all possible, avoid the practice of tip wiping. Technicians may do this out of habit, but there are very few valid reasons to wipe the tip.

Not Accounting For The Viscosity of A Sample

Image
Take, for example, a sample that contains large, sticky molecules – like glucose. The sample clings to the tip surface, traveling slowly when dispensed. Other less viscous and more volatile samples, such as ethanol, travel faster and have a tendency to evaporate. In both cases you must account for the physical propertie…
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Dispensing Liquids Too Quickly

  • Aliquoting bulk samples into single-use tubes is always a quick and easy task. The measured volume is set, and you are only working with a single tip, sample or mixture. But be mindful of pipetting a little too quickly: depending on the desired volume, the size of the tube, and where it is dispensed into the tube (such as at the upper wall or directed at the point at the very bottom) yo…
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Contamination by “Double Dipping”

  • On the bench, you might perform a variety of assays in which a reaction consists of multiple reagents – and your sample. You can introduce contamination by “double dipping:” when you use the same tip to draw and dispense a unique sample and then immediately pipette another sample without changing the tip first. Contamination like this can be tricky...
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Cleaning Irregularly—Or Not at All

  • Everyone knows what to do when liquid is aspirated entirely into the pipette barrel. That’s right, disassemble for a deep cleaning to prevent your pipette from becoming the source of contamination. But what about pipettes that pass through many hands without incident? Heavy use will cause parts to wear out more quickly, resulting in a significant decrease in how accurate…
See more on bitesizebio.com

Taking A Break only at The End of A Project

  • Whether you’re processing only a handful of samples, or hundreds at a time, pipetting is one of the most manually repetitive tasks that you perform in the laboratory. Some scientists take a project and power through from the first sample all the way to the last. Others practice pipetting equally between hands to try and reduce the amount of finger and hand strain, burn-out, and fatigue-ind…
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How Can You Avoid These Errors?

  • Many have learned how to pipette by measuring water samples into a tube on a scale, to see if the weight of the measured volume equals that of the set volume. Others have honed their skill by aliquoting a bulk sample into large numbers of individual tubes. So how can you help your laboratory mates to become pipetting experts and eliminate these top five errors? By giving amp…
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