Chloe Zhang
Filling a plastic pipette requires precision and care. Pipettes are commonly used in laboratories to measure and transfer specific volumes of fluids, and it is important to use them correctly to ensure accurate results. There are various types of pipettes, such as volumetric, Pasteur, and micropipettes, each with unique features and purposes. Understanding the different components of a pipette, such as the plunger, ejector, and volume adjustment wheel, is crucial for effective filling and dispensing. Proper filling techniques, including the use of pipette fillers, controllers, and guns, ensure accuracy, speed, and comfort during the filling process. In this discussion, we will delve into the step-by-step process of filling a plastic pipette, highlighting safety considerations and best practices to achieve precise and reproducible results.
| Characteristics | Values |
|---|---|
| Filling technique | Insert the plastic tip of the pipette just below the surface of the fluid and slowly release thumb pressure on the plunger to draw fluid into the pipette. |
| Transferring fluid | Transfer the pipette to the receiving vessel and place the tip just below the surface of the fluid. Depress the plunger slowly to expel the fluid. |
| Disposal | Do not use your mouth to draw fluid into the pipette. Follow lab guidelines for proper disposal of pipette tips. |
| Volume adjustment | Adjust the volume using the volume wheel on the side of the micropipette. |
| Calibration | Volumetric pipettes are calibrated for accuracy. When dispensing a volume, you can report it with two figures after the decimal point. |
| Tools | Pipette fillers, pipette controllers, and pipette guns are available to assist with filling and improve accuracy. |
| Compatibility | Some tools are compatible with plastic pipettes from 0.1 to 100 mL. |
| Speed | Certain tools offer multiple speed settings for filling and dispensing. |
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What You'll Learn
Prepare the pipette
To prepare a plastic pipette for use, you should first examine your micropipette. At the top, you will find a plunger that can be pushed in to empty the micropipette. Next to the plunger is an ejector that can be used to eject the plastic tip from the end of the micropipette. Along the side of the micropipette is a volume adjustment wheel that can be used to adjust the volume the pipette will take up or contain.
Look at the volume dial along the side of the micropipette. Micropipettes measure volumes in microliters. Determine what the volume is currently set to and adjust it with the volume adjustment wheel to reach the desired volume.
Insert the end of the micropipette shaft into one of the plastic tips in your plastic tip box. Place the rubber bulb (which looks a little like a turkey baster) at the top of the pipette and squeeze it to empty it of air. Remove the bulb and quickly cap the open top of the pipette with your finger. By tilting your finger to one side, allow a small amount of air into the pipette so the fluid drains out until the bottom of the meniscus (the curve-shaped depression in the fluid) reaches the fill mark or line.
Pipette fillers are also available to assist with filling plastic pipettes. These tools support manual pipetting and can improve accuracy and efficiency. Some pipette fillers offer speed and volume control, with fill and dispense speed settings for liquids of varying viscosities.
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Insert the pipette tip into the fluid
To insert the pipette tip into the fluid, start by examining your micropipette. At the top, you will find a plunger that can be pushed in to empty the micropipette. Next to the plunger is an ejector that can be used to eject the plastic tip from the end of the micropipette. Along the side of the micropipette, you will find a volume adjustment wheel that allows you to adjust the volume the pipette will take up or contain.
Look at the volume dial along the side of the micropipette to determine the current volume setting. Use the volume adjustment wheel to adjust the volume to the appropriate or desired level. Insert the end of the micropipette shaft into one of the plastic tips in your plastic tip box.
Now, insert the plastic tip of the pipette just below the surface of the fluid or water in your beaker. Slowly and gently release your thumb pressure on the plunger, drawing fluid into the plastic tip of the micropipette. Ensure that you do not submerge the pipette completely. Once the plunger has travelled all the way out, remove the pipette tip from the solution.
Transfer the pipette to the receiving vessel, which could be another beaker or a microfuge tube. Place the tip just below the surface of the fluid in the receiving vessel, maintaining the same level of submersion as before. Depress the plunger slowly and gently to expel all the fluid in the micropipette tip.
It is important to note that you should never draw fluid up into a Pasteur or volumetric pipette by sucking on it with your mouth. Always use the plunger mechanism to draw and expel fluid from the pipette.
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Draw fluid into the pipette
To draw fluid into a plastic pipette, start by inserting the plastic tip of the pipette just below the surface of the fluid in your beaker. Do not submerge it completely. Next, release your thumb pressure on the plunger—which should be located at the top of the micropipette—slowly and gently, drawing the fluid into the plastic tip.
If you are using a volumetric pipette, you will notice a number and a line or mark on the side. The number indicates the number of milliliters the pipette holds or dispenses when filled to the line or mark. Volumetric pipettes are calibrated to have a high level of accuracy, so you can report the volume dispensed with up to two figures after the decimal point. When you suck fluid into the pipette with the rubber bulb, the fluid level rises more slowly in the reservoir than in the tube above or below it.
Before drawing fluid into the pipette, you may need to adjust the volume the pipette will take up. To do this, locate the volume dial and adjustment wheel on the side of the micropipette. Determine the current volume setting and adjust as needed using the wheel.
There are also pipette fillers available to assist with filling plastic pipettes. These tools are designed to improve accuracy and comfort during manual pipetting tasks. Some pipette fillers offer speed and volume control, while others are rechargeable and offer multiple speed settings.
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Remove the pipette from the fluid
Once the plunger has travelled all the way out, remove the pipette from the solution. Transfer the pipette to the receiving vessel, beaker, or microfuge tube. Place the tip just below the surface of the fluid in the receiving vessel, but do not submerge it completely.
It is important to note that the removal process may vary depending on the type of pipette you are using. For example, with a micropipette, you would remove the pipette tip from the solution after expelling all the fluid.
If you are using a Pasteur pipette, you should refer to your lab's guidelines for specific instructions on how to work with and dispose of these items. Pasteur pipettes are often used as disposables, especially in biology labs, due to the risk of contamination with biological material.
In general, after removing the pipette from the fluid, you may need to follow specific protocols for disposing of the pipette tips or cleaning the pipette, depending on the type of pipette and the laboratory guidelines.
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Transfer the fluid to the receiving vessel
The reverse technique is used for solutions with a high viscosity or a tendency to foam. For reverse pipetting, press the operating button to the second stop. Dip the tip into the solution to a depth of 1 cm, and slowly release the operating button. This action will fill the tip with a volume that is larger than the set volume. Wait 1-2 seconds and withdraw the tip from the liquid, touching it against the edge of the reservoir to remove excess liquid. Dispense the liquid into the receiving vessel by pressing the operating button gently and steadily to the first stop.
For oily fluids, fill the pipette very slowly to prevent air bubbles. Wipe the tip against the edge of the vessel to remove excess liquid outside the tip before dispensing.
If you are using an automatic pipette, you can reverse pipette by pushing the plunger button all the way down and dispensing only to the first stop. Work quickly and deliberately over short distances.
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Frequently asked questions
First, insert the plastic tip of the pipette just below the fluid's surface in your beaker. Then, slowly release your thumb pressure on the plunger to draw the fluid into the tip. Once the plunger is fully extended, remove the tip from the solution.
Pipette guns are available with different functions, such as a rechargeable battery or a power source of electricity or compressed air. They are designed to reduce hand fatigue and improve accuracy.
Capp Controller is a lightweight and powerful instrument that can fill a 50ml pipette in less than 10 seconds. It has a lithium battery with no memory effect and can be charged while in use.
Volumetric pipettes have a number and a line or mark on the side. The number indicates the number of milliliters the pipette holds or dispenses when filled to the line. Use a rubber bulb to suck fluid into the pipette, and squeeze the bulb to empty it of air.
Pasteur pipettes are not designed to measure a specific volume. They are used to add drops or an indeterminate amount of a reagent. To use, simply follow your lab's guidelines on how to work with and dispose of them.
