Remove debris a nd lipids from th e sample whe n pipetting. Thermo scientific over 35 years of innovation thermo scientific pipetting guide a leader in pipetting for over 35 years we have led the way in liquid handling products and microplate instrumentation. A smooth rhythm will increase your throughput while. Techniques correct angle incorrect angle immersion angle keeping the immersion angle close to vertical can improve accuracy by up to 2. The pipetting cycle good pipetting technique youtube. Prewetting of the tip improves the analysis result. Advanced micropipette techniques for cell physiology. The selection of the correct tip, along with the right pipetting technique, can make these difficult solutions much easier to handle. The reverse technique is used for pipetting solutions with a high viscosity or a tendency to foam. Select an appropriate pipette for the volume you need to transfer all pipettes have a range in which they work, using a micropipette to.
This is a technique in which the plunger is depressed past the first stop to aspirate the aliquot from the sample, and depressed only to the first stop to deliver the aliquot. While mechanical pipettes are only suitable for forward and reverse pipetting, electronic dispensing systems frequently cover all of the above functions. A few physical factors such as altitude, temperature, and density of liquid can influence pipetting accuracyeven if your pipetting technique is perfectand that is something you should keep in mind. Good pipetting technique is essential for many biologists, but it can be hard to get right. Users guide en classic notice do not lubricate this pipette. Measurements, micropipetting, and sterile techniques laboratory l introduces. Manual stepper pipettes are ideal for simple repetitive tasks with a. Pipetting with an airdisplacement pipette is a five step process that is often referred to as forward pipetting. The department of biological sciences at the university of alberta brings you this video tutorial specifically relevant to your student laboratory courses, specifically microbiology. Dip the tip into the solution to a depth of 1 cm, and slowly release the operating button. We have always ensured that innovation, ergonomics, accuracy, precision and safety are key aspects of our products designs. Use of the reverse pipetting technique can improve the precision when pipetting such solutions, since it aspirates a volume that is greater than that set.
Slow aspiration allows the liquid to rise to an equilibrium level and slow dispensing reduces the effect of the liquid retention film. This is a technique in which the plunger is depressed past the first stop to aspirate the aliquot from the sample, and. Select an appropriate pipette for the volume you need to transfer all pipettes have a range in which they work, using a micropipette to transfer volumes outside that range can break the pipette pipettes will have their range written somewhere on them ex. Get a printable copy pdf file of the complete article 311k, or click on a page image below to browse page by page. As part of our good pipetting practice program, mettler toledo has developed a. Small deviations in the volumes associated with cell. Northwest association for biomedical research updated july 19, 2012 5 12. Good pipetting practice pipetting calibration and technique. Pipetting calibration and technique how they affect. Oct 23, 2014 pipetting with an airdisplacement pipette is a five step process that is often referred to as forward pipetting. Impact of pipettingtechnique t he pipette is eliminating sources. Use a consistent pipetting rhythm from sample to sample. Improve pipetting performance with pipette systems. Place the end of the tip against the inside wall of the recipient vessel at an angle of 10 to 40.
Utilizing the hydrophobicselfassembledmonolayer films patterned in microchannels as pressurecontrolled valves, a small. The technique is mainly recommended for solutions with a high viscosity or a tendency to. A picoliter pipetting technique using the microfluidic method is presented. Pipetting and dilutions pages 1 3 text version fliphtml5.
Reverse pipetting technique the reverse technique is used for pipetting solutions with a high viscosity or a tendency to foam. Forward pipetting is a technique to dispense a measured quantity of liquid by means of air displacement pipette. Wipe the tip against the edge of the vessel to remove excess liquid outside the tip before dispensing. Obviously, this is not the case, since many factors associated with pipettes. If you are inexperienced with this procedure or unsure whether pipetting is a suitable transfer method consult notes in lab manual and obtain instruction from experienced laboratory personal before.
Reliable accuracy and precision, increased efficiency, uncompromised safety, advanced ergonomics all are the hallmarks of innovative product design. Pdf controllable picoliter pipetting using hydrophobic. Pipetting procedures the most important aspect of pipette calibration is being consistent in your procedure and having a good cadence. Air displacement pipetting air displacement pipetting is highly accurate for standard pipetting applications. In case of solutions with a high viscosity or a tendency to foam, reverse pipetting is more suitable. While mechanical pipettes are only suitable for forward and reverse pipetting, electronic dispensing systems frequently. How you pipette directly affects the success and repeatability of your experiments. With viscous or foaming liquids or when dispensing very small sample volumes the results may be greatly improved by reverse pipetting. These five simple techniques alone can have a big impact. Increasing precision when pipetting protein samples. Holding the pipette at an angle as it is removed from the sample alters the volume aspirated.
Use these good pipetting practicetm techniques to improve your pipetting performance. Two main techniques exist and each should be applied depending on the sample liquid. Good laboratory pipetting thermo fisher scientific. Pipetting techniques pdf 8 tips to improve your pipetting technique. Laboratory solutions from mettler toledo 29,490 views. How to use pipetting techniques in the lab science. Full text full text is available as a scanned copy of the original print version.
The technique is mainly recommended for aqueous solutions, such as buffers, or diluted acids or alkalis. Dip the tip into the solution to a depth of 1 cm, and slowly. The results can be improved by reducing the pipetting speed. The technique of pipetting at a 45 to 60 degree angle is appropriate for sterile technique pipetting associated most commonly with cell culture.
This technique is used for pipetting heterogeneous samples, such as blood or serum. This technique is especially important when pipetting small volumes file of the complete article 311k, or click on a page image below to browse page by page. Aspire and expel an amount of the sample liquid at least 3 times before aspirating a sample for delivery. Thermo scientific pipetting guide recommendations for pipetting different compounds solution compound examples pipette tip pipetting technique comments aqueous solution buffers, diluted salt solutions air displacement standard forward viscous solution protein and nucleic acid solutions, glycerol, tween 20406080 air displacement positive. Wait for at least a second, then press the push button to the second stop to expel. Follow these ergonomics suggestions while pipetting. Push the piston down to the purge position the second stop, then draw the liquid up. Reverse mode pipetting is a method of pipetting commonly used with viscous liquids. However, conditions, such as atmospheric pressure as well as the specific gravity and viscosity of the solution, may effect the performance of air displacement pipettes. Reverse pipetting is only possible with air displacement pipettes. Residual serum can sometimes be found on the outer surface of the tip. Use standard mode pipetting choose standard mode pipetting rather than reverse mode, for all but viscous samples, if accurate and precise results are desired. Utilizing polymer technology makes the inner surface of the pipette tip more.
Touching the sides of the container can cause wicking and a loss of volume due to the effects of surface tension. Especially when pipetting small volumes the influence of the pipetting technique can have tremendous effects on the experimental result. Selection of the dispensing technique suitable in each case may have a significant impact on the analysis. To attach, firmly press the shaft of the pipette into the large open end of the tip with light force. In this short video, we demonstrate the five distinct steps of pipetting and how. Especially when pipetting small volumes the influence of the pipetting. Pipetting techniques rainin gpp good pipetting practice. To attach, firmly press the shaft of the pipette into the large open end of the tip with light force to ensure a good seal. Reverse pipetting is a technique to dispense a measured quantity of liquid by means of air displacement pipette. Refer to the test insert to determin e if skim and or whole milk samples c an be used as sa mples. With regards to pipetting of, for example, viscous liquids, mainly slow pipetting speed, as well as reverse pipetting, are applied. Reverse pipetting should be used if high accuracy is needed. Use this technique when pipetting viscous liquids or volatile solvents.
If greater precision is needed, a serological pipette may be used in combination with a micropipettor. Training tips are homemade sets of marked pipette tips that guide the trainee as they learn to use micropipettes. Reverse mode pipetting can make obtaining accurate and precise results for the pipetting. Make sure the tip is sufficiently below the surface. There is nothing wrong with that approachthey are, indeed, highly accurate and used for standard pipetting applications. Pipetting pipetting pipetman cl assic pipetman cl assic users guide 9 dispense 1. Dec 30, 2010 the pipetting cycle good pipetting technique duration. Finnpipette f2 pipettes and finntip flex pipette tips pdf, 229kb in this application note we test the pipetting performance of. Most liquid types can be dispensed using forward pipetting table 1. Impact of pipetting techniques on precision and accuracy. Introduction to micropipetting the micropipette is a device used to measure and deliver microliter ul quantities of liquid. Below is a demonstration of pipetting results obtained by the same operator, performing repetitive pipetting of 10 aliquots of 20. A few physical factors such as altitude, temperature, and density of liquid can.
Press the push button slowly and smoothly to the first stop. Do not use an adjustable pipette outside its recommended range. Pipetting 3600 how you pipette directly affects the success and repeatability of your experiments. The technique is mainly recommended for solutions with a high viscosity or a tendency to foam. L, with electronic pipettes from different manufacturers. Pipetting technique guide transferring liquid with a micropipette 1. Do not repetitively bang the pipette shaft into the tip.
Small deviations in the volumes associated with cell culture are not critical to the success of cell growth making the sterile technique preferable in that situation. Drain a pipette with tip against the inner wall of the. Impact of pipettingtechnique t he pipette is eliminating. The pipetting cycle good pipetting technique duration. These five simple techniques alone can have a big impact on your results, but there are a few more to consider. The following aqueous pipetting data headl head8, was acquired from eight 8 new quadra4 450ul heads. Choosing the correct pipetting technique helps securing the accuracy and precision necessary for reproducible, reliable results. To ensure precise and accurate pipetting results volume in. All testing was done at 230c 50% relative humidity. The forward mode is the standard way of pipetting with an air displacement pipette like pipetman. When i take 1 l of liquid using a micropipette, i seem to always take less than 1 l, and that amount is variableinconsistent. The technique is mainly recommended for aqueous solutions, such as buffers, or diluted acids.