Thermo Scientific Pipetting Guide

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Tips for Good Laboratory Pipetting

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Chemical compatibility of plastics Customer support shooting Trouble Contents Introduction Pipetting terms Pipetting of pipettes Types General guidelines pipetting techniques Pipetting Tip information compounds different Recommendations for pipetting Ensuring optimum performance Usage of Novus Finpipette the accuracy Factors of affecting air displacement pipettes cross-contamination Preventing Maintenance of your Finnpipettes Autoclaving UV resistance Calibrating (incl. your table)pipettes conversion On-line calibration pipette General guidelines for decontaminating pipettes

Thermo Scientific Pipetting Guide Thermo Scientific Over 35 Years of Innovation

A leader in pipetting

For over 35 years we have led the way in liquid handling products and microplate instrumentation. We have always ensured that innovation, ergonomics, accuracy, precision and safety are key aspects of our products’ designs. In 1971 we introduced Thermo Scientific Finnpipettes, the world’s first continuously variable micropi- pettes. In 1976 we introduced the world’s first multichannel pipette. Since then we have continuously improved our pipettes, always leading the way with ergonomic design. Over the last 35 years, innovations such as the improved finger rest, soft-touch tip ejection and super blow-out have made Finnpipettes increasingly user-friendly. Our intensive research program and commitment to our customers forms the foundation for future innovations. The new demands in pipette applications are the key drivers of our product development. To date, over 3 mil- lion Finnpipettes have been sold in 150 countries. Thanks to our extensive R&D and feedback from the field, Thermo Scientific Pipetting Guide Finnpipettes are the preferred choice for optimal liquid handling results, with our multichannel pipettes being the market leader worldwide.

Most clinical and research laboratories are equipped with manual or electronic pipettes for dispensing liquids. The amount of liquid can range from ml’s to below 1 μl, especially when using expensive reagents. In all cases, the dispensing must be accurate and precise to guarantee good research results. Not only the pipette but also the tip plays a crucial role as these two form together the pipetting system.

As pipetting is often a repetitive task, the pipettes must be designed so that the risk of hand and upper limb stress is minimized.

We offer a wide range of high-quality manual and electronic pipettes and pipette tips to ensure good laboratory pipetting. In this guide we will outline some practical aspects of pipetting to help you to get the best possible results.

Pipetting terms

The following terms are used throughout this guide and in instructions for use. We will explain them briefly:

Aspirate – to draw up the liquid into the pipette tip

Dispense – to discharge the liquid from the tip

Blow-out – to discharge the residual liquid from the tip

Calibration check – to check the difference between the dispensed liquid and the selected volume

Adjustment – altering the pipette so that the dispensed volume is within the specifications

- When the operating is button After immersing After the tip into the specified volume of liquid is aspirated into the tip. 4. pressed to the first stop again, the empty To air dispenses the liquid. the tip completely the operating is button pressed to the second stop (blow out). 3. the liquid, the operating is button released. This creates a partial vacuum and -

When the operating is button The The piston to moves the appropri 2. pressed to the first stop, the piston expels the same volume of air as indicated on the volume setting. How How does an air displacement pipette work? 1. ate position when the volume is set. - pipette pipette tip piston an effect on the an performance effect of air displacement pipettes. Air displacement used pipetting, for standard applications, pipetting condi is highly accurate. However, tions as such atmospheric pressure as well as the specific gravity and viscosity of the solution may have Air displacement pipetting Air displacement Both Both types a of have piston pipettes that in moves a cylinder or in a In capillary. air displacement a pipettes, cer tain volume of air remains between the piston and the liquid. In positive displacement the pipetting, piston is in direct contact with the liquid. Air displacement are pipettes meant for general use with aqueous displacement are solutions. pipettes Positive used for high viscosity and volatile liquids. Types of pipettes Types There are two types of air pipettes: displacement and positive displacement pipettes.

Thermo Scientific Pipetting Guide Types of pipettes Types of pipettes Types

Positive displacement pipetting How does Thermo Scientific Pipetting Guide Finnpipette Stepper work? The repeater pipette Thermo 50 ml 50 Scientific Finnpipette® Stepper uses 1. The piston inside the tip moves ®

the positive displacement principle. up when filling the tip with liquid. Finntip Stepper microsyringe tips have a piston inside a cylinder unit; these 2. When the dispensing lever is tips are disposable. This helps to pressed down, the piston moves 1=1 ml 1=1 avoid sample to-sample cross-con- down and the selected volume is

tamination (also known as sample dispensed. The dispensing lever has Thermo Scientific carry-over), and contamination due to be pressed once for each dis- to the aerosol effect. pensing stroke (= step).

Finnpipette Focus Finnpipette Stepper

Gnerealguidelines pipetting ScientificThermo

General pipetting guidelines

Check your pipette at the beginning • Pre-rinsing (three - five times) the • Avoid contamination to or from Pipetting Guide Pipetting of your working day for dust and tip with the liquid to be pipetted fingers by using the tip ejector. dirt on the outside. If needed, wipe improves accuracy. This is especially with 70% ethanol. important when pipetting volatile • Always store pipettes in an compounds since it prevents liquid upright position when not in use. • Check that you are using tips rec- dropping out of the tip. Finnpipette stands are ideal for this ommended by the manufacturer. To purpose. ensure accuracy, use only high-qual- • Pipette parallel samples in a simi- ity tips made from contamination- lar way. free polypropylene. • Avoid turning the pipette on its • Tips are designed for single use. side when there is liquid in the tip. They should not be cleaned for Liquid might go to the interior of reuse as their metrological charac- the pipette and contaminate the teristics will no longer be reliable. pipette.

Forward pipetting

Ready position 1 2 3 4 Otherwise no: formation of bubbles 3. Dispense the liquid into the or foam in the tip or in the test tube receiving vessel by gently press- First stop or well. ing the operating button to the first Second stop stop. After one second, press the 1. Press the operating button to the operating button to the second first stop. stop. This action will empty the tip. Yes: when pipetting and mixing Remove the tip from the vessel, a sample or reagent into another 2. Dip the tip into the solution to a sliding it along the wall of the liquid. depth of 1 cm, and slowly release vessel. the operating button. Withdraw The forward technique is recom- the tip from the liquid, touching it 4. Release the operating button to mended for aqueous solutions like against the edge of the reservoir to the ready position. buffers, diluted acids or alkalis. remove excess liquid.

Repetitive pipetting

Ready position 1 2 3 4 1. Press the operating button to the 3. Dispense the liquid into the second stop. receiving vessel by gently press- First stop ing the operating button to the first Second stop 2. Dip the tip into the solution to a stop. Hold the button in this posi- depth of 1 cm, and slowly release tion. Some liquid will remain in the the operating button. Withdraw Yes: especially for adding of rea- tip, and this should not be dis- the tip from the liquid, touching it gents into tubes or into the wells of pensed. against the edge of the reservoir to a microtiter plate. remove excess liquid. 4. Continue pipetting by repeating This technique is intended for steps 2 and 3. repeated pipetting of the same volume.

Reverse pipetting viscosity or a tendency to foam. 3. Dispense the liquid into the Ready position 1 2 3 4 5 This method is also recommended receiving vessel by depressing the First stop for dispensing small volumes. It is operating button gently and steadily Pipetting Guide only possible with air displacement

Second stop to the first stop. Hold the button pipetting General guidelines pipettes. in this position. Some liquid will Yes: for pipetting of samples or 1. Press the operating button to the remain in the tip, and this should reagents when no mixing into second stop. not be dispensed. another liquid is required. 2. Dip the tip into the solution to a 4. The liquid remaining in the tip depth of 1 cm, and slowly release can be pipetted back into the origi- Reverse pipetting avoids the risk of the operating button. This action will nal solution or thrown away with splashing, foam or bubble formation. fill the tip. Withdraw the tip from the tip. Thermo Scientific Thermo Scientific the liquid, touching it against the The reverse technique is used for 5. Release the operating button to edge of the reservoir to remove pipetting solutions with a high the ready position. excess liquid.

Pipetting of heterogeneous samples

1. 3. + 4. Press the operating button to Ready position 1 2 3 4 5 6 Press the operating button to the first stop. Dip the tip into the sam- the first stop and release slowly to First stop ple. Make sure the tip is sufficiently the ready position. Do not remove Second stop below the surface. the tip from the solution. Repeat this process until the interior wall of 2. Release the operating but- the tip is clear. Yes: when prerinsing the tip is not ton slowly to the ready position. possible and the full sample should This action will fill the tip with the 5. Remove the tip from the solution be dispensed for correct analysis. sample. Remove the tip from the by sliding it along the wall of the vessel. Press the operating button This technique is used for pipetting solution by sliding it along the wall to the second stop and completely heterogeneous samples like blood of the vessel. Dip the tip into the empty the tip. or serum. target solution. Make sure the tip is sufficiently below the surface. 6. Release the operating button to the ready position.

Tip information

Pipette tips are part of the complete Thanks to the unique sealing- The Finntip® range covers basic and pipetting system, and the quality end design, Finntips form a leak special tips, including wide orifice of both pipette and tip are impor- proof seal with the tip cone of the tips, stepper tips and other special tant. Thermo Scientific Finntips are pipette. This prevents leakage and models and so offers a suitable tip designed using our knowledge of guarantees optimum accuracy and for most applications. liquid handling and are made of precision. The surfaces of Finntips high-quality raw materials. are uniformly smooth and hydropho- bic. This minimizes surface wetting and allows complete dispensing of the liquid.

Recommendations for pipetting differentRecommendationscompoundspipetting for ScientificThermo

Recommendations for pipetting different compounds

Solution/ Examples Pipette Tip Pipetting Comments compound technique Pipetting Guide Pipetting Aqueous Buffers, diluted salt Air displacement Standard Forward solution solutions

Viscous Protein and nucleic Air displacement Standard or wide orfice Reverse Pipette slowly to avoid bubble formation. solution acid solutions, glycerol, Tween 20/40/60/80 Positive displacement Positive displacement

Volatile Methanol, hexane Air displacement Filter Reverse Pipette rapidly to avoid evaporation. Carbon compounds filter tips prevent vapor going into the pipette Positive displacement Positive displacement very effectively

Body fluids whole blood, serum Air displacement Standard or wide Pipetting Residual liquid can be found on the outer orifice tip of hetero- surface of the tip. Wipe the tip against the geneous edge of the vessel to remove this liquid samples before dispensing.

Nucleotide Gonomic DNA, PCR Air displacement Filter or wide orifice Forward For genomic DNA wide orifice tips can be solutions products used to eliminate mechanical shearing. Positive displacement Positive displacement

Radioactive 14Carbonate, 3H-thy- Air displacement Filter Forward compounds midine Positive displacement Positive displacement

Acid/alkalis H2SO4, HCI, NaOH Air displacement Filter Forward Toxic Air displacement Filter Forward or samples reverse Positive displacement Positive displacement

Ensuring Optimum Performance

Error‑free pipetting requires both precision and accuracy. A number of factors can affect these specifications. These form the main quantitative parameters for evaluating pipette performance.

What are accuracy and precision? For example when the set volume is 20 µl:

Accurate, but not precise: The mean

volume is the correct (set) volume, 20µl 20µl 20µl 20µl 20µl Accurate, but but the separate pipettings differ 20µl 20µl 20µl 20µl 20µl not precise from the set volume. 20µl 20µl 20µl 20µl 20µl

Precise, but not accurate: There is 20µl 20µl 20µl 20µl 20µl no variation between the separate 20µl 20µl 20µl 20µl 20µl Precise, but pipettings,but the mean volume not accurate differs from the set volume. 20µl 20µl 20µl 20µl 20µl

20µl 20µl 20µl 20µl 20µl

Accurate and precise: The mean 20µl 20µl 20µl 20µl 20µl volume is the set volume and there Accurate 20µl 20µl 20µl 20µl 20µl is no variation between the and precise different pipettings.

Usage of Thermo Scientific Finnpipette Novus

Finnpipette Novus is an electronically assisted pipette for a wide range of liquid handling operations. Pipetting Guide

Thanks to the electronic motor and electronic control, pipetting is easy and comfortable, yet still fast and of Finnpipette Usage Novus accurate. It operates on the air displacement principle and uses detachable, disposable tips, which are easy to remove with a soft touch tip ejector.

The guiding and easy user-interface is very fast to learn. The adjusted delivery volume is clearly indicated in the LCD display on top of the handle. The long lasting Lithium-Ion -battery is always charged with rapid charge technique. If needed, the battery can be charged over the lunch hour.

The adjustable, index finger operated pipetting trigger uses natural hand movement, increasing comfort and reducing the risk of repetitive stress injuries. Thermo Scientific Thermo Scientific The menu map of Finnpipette Novus is shown below.

PIPETTE RPIPET STEPPER DILUTE PROG 1 PROGRAM PROG 2 OPTIONS . . . PROG 9 PIPETTE MIX+PIPETTE PIPETTE+COUNT RPIPET CALIBRATE STEPPER SERVICE SEQ+STEPPER NAME DILUTE POWER TWO POINTS MIX+DILUTE BUZZER ONE POINT MANUAL VERSION COUNTER SEQ+ASPIRATE

PIPETTE = forward pipetting

RPIPET = reverse pipetting or repetitive pipetting

STEPPER = repeated dispensing of a selected volume. The minimum stepper volume is 5% of the maximum volume e.g. for FP Novus 1-10 µl the minimum stepper volume 0.5 µl.

DILUTE = dispensing of two selected volumes (between the first and the second liquid an air buffer is aspirated)

PROGRAM

Programs are stored settings that can be edited, stored and retrieved. In the program mode additional functions are available:

MIX+PIPETTE = this function adds automatic mixing after normal pipetting

PIPETTE+COUNT = this function adds automatic count number to pipetting

SEQ+STEPPER = the sequential stepper mode enables serial dispensing of different volumes (opposite to the normal stepper mode where only one fixed volume can be dispensed).

MIX+DILUTE = this function adds automatic mixing after diluting

MANUAL = in this mode it is possible to measure volumes. In the manual mode only slower speeds are available to ensure a quick stop. The slow pipetting speed is useful e.g. when applying fixative to cells where a regular speed would damage the cells. The manual mode can also be used for titration.

SEQ+ASPIRATE = the sequential aspirate mode enables serial aspirating of different volumes. -

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3 1.2% - 1.2% Accuracy 1

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Finnpipet 3 ®

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te Finnpipet ® 1. Pipette held vertically, held Pipette vertically, 1. tip immersed about 1 cm into the liquid. boiling point decreases quite close to room will temperature, which increase the evaporation loss dramatically. temperature. Typically the density for water Typically is 0.998 kg/dm temperature. sulfuric acid (95-98% H Altitude the The geographic accuracy The altitude air through affects air pressure pressure. decreases in higher altitudes and the factor Z conversion decreases as well. Also, with some liquids the Density Density The density the (mass/volume liquid ratio) volume affects that is aspirated into the tip. A small er dose of liquid with higher density than water is aspirated compared to similar operation With densityThis lower is is caused liquids the by the with the opposite. effect water. flexible dead The densityair volume along of with liquids the varies also earth according gravity. to the Temperature Temperature 1 cm Pipetting Pipetting position (e.g. when using a 1-10 ml pipette) Temperature has many effects on pipetting accuracy. The The factor that has the greatest effect on accuracy. has pipetting many effects Temperature is between the the temperature used difference The air delivery gap device (dead and liquid. air volume) between the liquid surface and the piston experiences thermal expansion effects This either according to reduces or the increases case. the liquid amount aspirated into the tip along with other effects. Factors Affecting of Pipettes the Air Factors Accuracy Displacement 10

Thermo Scientific Pipetting Guide Factors Affecting the Accuracy of Air Displacement Pipettes Preventing cross-contamination Pipetting Pipetting Guide Preventing cross-contamination Preventing Preventing cross-contamination Preventing

Pipette-to-sample Sample-to-pipette Sample-to-sample (carry-over)

A contaminated pipette or contami- Samples or aerosols from samples The remains of sample A can mix nated tips can cause contamination can enter the cone of the pipette. with next sample B inside the tip of samples. and may cause a false test result. Thermo Scientific Thermo Scientific Pipetting Guide

Prevention: Prevention: Prevention:

• Use sterilised tips or sterilised • Keep the pipette vertical when • Change the tip after each sample. filter tips and if possible autoclave pipetting in order to prevent liquid the pipette. from running into the pipette body. • If you suspect that your pipette is contaminated, autoclave or clean • Change the tip after pipetting of • Release the push-button slowly. your pipette (see “Maintenance of each sample. your Finnpipette” and “Autoclaving”, • To avoid aerosol contamination, use filter tips or use a positive displacement pipette and tips.

• Store the pipette vertically.

11 - - 3. After autoclaving, 3. After the pipette must be cooled at room tempera ture for at least two hours before use. 4. the Check calibration every after treatment. autoclave Please see the Instructions for Use manual for maintenance instruc can find the latest You version tions. of the manuals from www.thermo. com/finnpipette. 21°C 0 minutes. The The tip cone 20 minutes. . All Finnpipette Focus and 2. All Focus DigitalFinnpipette models can be (steam autoclaved sterilized) in one piece at 1 (248°F) for modules of Novus and Finnpipette BioControl Finnpipette can also be autoclaved. • Short-term service: If the pipette is it used is daily, recommended to the calibration every check three months. • Long-term service: If the pipette is used daily it should be serviced every six months. -

20 minutes. 21°C (248°F) for Autoclaving UV UV resistance and Focus DigitalFinnpipette might models colour The handle change are from however UV grey to resistant.light If the inner parts yellow. of the are pipette exposed to UV that light, there please is grease check sufficient on the piston and O-rings. the tips are moist. Allow moisture to before using evaporate the tips, overnight. preferably Please, follow Please, these follow instructions carefully in order to damage avoid to tips and pipettes. (steam sterilize) tips Autoclave 1. at 1 Immediately after Immediately autoclaving, after be checked for dust and dirt be on checked the outside surfaces. atten Particular tion should be paid to the tip cone. No other 70% solvents except ethanol should be used to clean the pipette. come with detailed instructions on come with detailed instructions to disassemble the pipette. how in-lab routine for Instructions included. maintenance are also At the • beginning Daily checking: the should pipette of workday, each Maintenance of your Thermo Scientific Thermo Scientific Finnpipette of your Maintenance easy to service are Finnpipettes Finnpipettes All in the laboratory. 2 1

Thermo Scientific Pipetiing Guide Maitenance of your Finnpipette Calibration of Pipettes Calibration of Pipettes Calibration

All Finnpipettes are factory calibrated and adjusted to give the volumes as specified with Pipetting Guide water. During factory calibration, the performance is checked with 5 weighings at both the minimum and maximum volumes of the volume range. A calibration report is included with every pipette. Normally, the pipettes do not need adjustment, but they are constructed to per- mit re-adjustment for liquids of different temperature and viscosity.

Calibration of pipettes means deter- standards e.g. EN ISO 8655 mul- the maximum selectable volume. mining the difference between the tiplied by two. The actual standard The μl limit of the nominal volume dispensed volume and the selected specifications, and if the highest applies to every selectable volume Thermo Scientific volume. Adjustment means altering accuracy is needed, the manufac- throughout the volume range. For the pipette so that the dispensed turer’s specifications, should be example, for a 10-100 μl pipette the volume is within the specifications. used only when testing can be per- maximum permissible accuracy formed in a controlled environment limit is 0.8 μl and the maximum Calibration of pipettes in a using distilled or deionised water. permissible precision limit is 0.3 μl. quality system With multichannel pipettes these Device requirements and test values are further doubled. The main objective of pipette conditions calibration in a quality system is to The EN ISO 8655 specifications are ensure that dispensing is carried An analytical balance must be used. shown in the table below. out with the intended accuracy. The scale graduation value of the Very often the error limits are taken balance should be chosen according Procedure to check calibration from the manufacturer’s specifica- to the selected pipette volume. tions, although far less accuracy The pipette is checked with the is needed to perform the task. It Pipette specifications according maximum volume (nominal volume) should be kept in mind that in a lab- to EN ISO 8655 and with the minimum volume oratory environment (uncontrolled) or 10% of the maximum volume, the manufacturer´s specifications The EN ISO 8655 standard gives whichever is higher. For example, may not be achieved. Therefore the accuracy and precision limits as Finnpipette 0.5-10 µl is tested at 10 every user should define their own both absolute and relative values. µl and 1 µl. A new tip is first pre- acceptance limits, according to the The values are specified for fixed wetted 3-5 times and a series application and the ambient condi- single channel air displacement tions. Another option is to use the pipettes. With variable volume acceptance limits stated in the pipettes, the nominal volume is

Volume range Readability Weighing Sartorius model Article description (mg) capacity (g) number (example) under 10 µl 0.001 5.1 MC5 Standard version under 10 µl 0.001 5.1 MC5 / VF988 Special version with special draft shield and humidity trap 10 – 100 µl 0.01 210 ME215S Standard version 10 – 100 µl 0.01 210 ME215S / VF2397 Special version with special draft shield above 100 µl 0.01 / 0.02 / 0.05 60 / 110 / 210 ME215P Standard version

13 = 1...n)

2 v

– v) i n – 1 (v n  i=1

= Single measurement result (i i n = Number of measurements v = Mean volume v Precision (random error) Precision to Precision refers the repeatability of the It pipettings. is expressed as standard deviation (s) or coefficient of variation (cv). In addition to the of the laboratoryfeatures pipette, practice and user experience are the precision. main factors affecting s = s = Standard deviation Standard deviation can be expressed as a relative value as cv. cv = x 100% s/ - - 2 mg.

µl 0,05 0,04 0,075 0,08 0,1 0,2 0,3 0,6 1,5 3,0 6,0 15,0 30,0 0 V = Mean volume

% 5,0 2,0 1,5 0,8 0,5 0,4 0,3 0,3 0,3 0,3 0,3 0,3 0,3 Maximum Maximum permissible random error (CV) 0 = Nominal volume 0 Accuracy Accuracy can be expressed as a relative value: A% V = x A 100% / seconds and the mass loss is If an trap evaporation or lid on the vessel is used, an cor evaporation rection is unnecessary. factor Z Conversion is for calculat ing the density of water suspended in air at a test temperature and pressure. See the tableconversion on page 74. error) (systematic Accuracy Accuracy between is the difference the dispensed volume and the selected volume of a pipette. V A -V = A = Accuracy V

- ±µl 0,05 0,08 0,125 0,12 0,2 0,5 0,8 1,6 4,0 8,0 16,0 40,0 60,0 -

±% 5,0 4,0 2,5 1,2 1,0 1,0 0,8 0,8 0,8 0,8 0,8 0,8 0,6 Maximum permissible Maximum permissible error (ACC) systematic w = Weight (mg) Weight = w

Nominal Nominal ISO 8655 pipettes air error limits for single displacement channel µl 1 2 5 10 20 50 100 200 500 1000 2000 5000 10000 volume ing and begin timing with a stop the how much read Check watch. ing decreases during 30 seconds. Compare this to the pipetting. time pipetting might be 10 Typically, Z = factor for Conversion mg/µl conversion Evaporation loss can be significant determine To with low volumes. mass loss, dispense water into the weighing vessel, note the read Formulas for calculating results results for calculating Formulas of Conversion mass to volume V = (w + e) x Z (µl) V Volume = A pipette is always A adjusted is pipette for always delivery (EX) of the selected vol ume. If the calculated results are within the selected limits, the adjustment of the is pipette correct. of ten is pipettings performed with With both multichannel volumes. both pipettes, volumes are tested with the two edge channels. e = Evaporation loss (mg) 14

Thermo Scientific Pipetting Guide Calibration Conversion Table Table Conversion

Values of the conversion factor Z (µl/mg), as a function of temperature and Pipetting Guide pressure, for distilled water.

Temperature °C Air pressure kPa* 80 85 90 95 100 101 105 15,00 1,0017 1,0018 1,0019 1,0019 1,0020 1,0020 1,0020 15,50 1,0018 1,0019 1,0019 1,0020 1,0020 1,0020 1,0021 16,00 1,0019 1,0020 1,0020 1,0021 1,0021 1,0021 1,0022 16,50 1,0020 1,0020 1,0021 1,0021 1,0022 1,0022 1,0022 17,00 1,0021 1,0021 1,0022 1,0022 1,0023 1,0023 1,0023

17,50 1,0022 1,0022 1,0023 1,0023 1,0024 1,0024 1,0024 Thermo Scientific 18,00 1,0022 1,0023 1,0023 1,0024 1,0025 1,0025 1,0025 18,50 1,0023 1,0024 1,0024 1,0025 1,0025 1,0026 1,0026 19,00 1,0024 1,0025 1,0025 1,0026 1,0026 1,0027 1,0027 19,50 1,0025 1,0026 1,0026 1,0027 1,0027 1,0028 1,0028 20,00 1,0026 1,0027 1,0027 1,0028 1,0028 1,0029 1,0029 20,50 1,0027 1,0028 1,0028 1,0029 1,0029 1,0030 1,0030 21,00 1,0028 1,0029 1,0029 1,0030 1,0031 1,0031 1,0031 21,50 1,0030 1,0030 1,0031 1,0031 1,0032 1,0032 1,0032 22,00 1,0031 1,0031 1,0032 1,0032 1,0033 1,0033 1,0033 22,50 1,0032 1,0032 1,0033 1,0033 1,0034 1,0034 1,0034 23,00 1,0033 1,0033 1,0034 1,0034 1,0035 1,0035 1,0036 23,50 1,0034 1,0035 1,0035 1,0036 1,0036 1,0036 1,0037 24,00 1,0035 1,0036 1,0036 1,0037 1,0037 1,0038 1,0038 24,50 1,0037 1,0037 1,0038 1,0038 1,0039 1,0039 1,0039 25,00 1,0038 1,0038 1,0039 1,0039 1,0040 1,0040 1,0040 25,50 1,0039 1,0040 1,0040 1,0041 1,0041 1,0041 1,0042 26,00 1,0040 1,0041 1,0041 1,0042 1,0042 1,0043 1,0043 26,50 1,0042 1,0042 1,0043 1,0043 1,0044 1,0044 1,0044 27,00 1,0043 1,0044 1,0044 1,0045 1,0045 1,0045 1,0046 27,50 1,0045 1,0045 1,0046 1,0046 1,0047 1,0047 1,0047 28,00 1,0046 1,0046 1,0047 1,0047 1,0048 1,0048 1,0048 28,50 1,0047 1,0048 1,0048 1,0049 1,0049 1,0050 1,0050 29,00 1,0049 1,0049 1,0050 1,0050 1,0051 1,0051 1,0051 29,50 1,0050 1,0051 1,0051 1,0052 1,0052 1,0052 1,0053 30,00 1,0052 1,0052 1,0053 1,0053 1,0054 1,0054 1,0054

*1kPa = 10 hPa

On-line pipette calibration

We offer an easy-to-use pipette lower calibration volume and ten the “Calculate” button to calculate calibration calculator at www. pipettings with the nominal volume. the accuracy and precision. If the thermo.com/finnpipette. This can be According to ISO 8655, with mul- results are within the limits, the found from the Finnpipette product tichannel pipettes both volumes adjustment of the pipette is correct. pages or by using the search option should be tested with all channels. If not, the pipette has to be read- pipette calibration. Procedures how Enter the weighing results into the justed with the lower volume and to check the calibration and how fields. For multichannel pipettes, checked again. to adjust the pipette are included. the channel number can be entered Briefly, do ten pipettings with the into the serial number field. Press

spores can spores be by inactivated wiping the aerosols from aerosols the can acids enter the aerosols from aerosols the can enter alkalis the autoclave it according to it autoclave the according manufac- “potentially infectious liquids”. “potentially The is normally evaporation process* grease that comes with that grease comes the pipette. used The in plastics Finnpipettes The tip cone and tip ejector can be are acid resistant. However, are However, acid resistant. are However, alkali resistant. lower lower part of the pipette and affect the the pipette with a moistened tissue with performance of performance the pipette. as Clean described above in “Aqueous and solutions buffers”. used The in plastics Finnpipettes contaminated parts contaminated in detergent. Rinse cleaning solutions. cleaning Rinse with well and dry at water maximum 60°C in a dryerdistilled using the compartment. grease piston Lubricate if neccessary with that comes the pipette. Open pipette, rinse pipette with Autoclave according to instructions. Viruses and Viruses to instructions. according Autoclave at at maximum 60°C in a dryer compartment. pressure. Alternatively, immerse the Alternatively, pressure. compartment. Lubricate piston if neccessary using the compartment. piston grease Lubricate if neccessary with that comes the pipette. If you have an pipette, autoclavable lower lower part of the pipette and affect the of performance the pipette. as Clean described above in “Aqueous solutions and buffers”. Use 70% to ethanol disinfect. left left in an bath ethanol overnight. sufficient for sufficient liquids with a high vapour parts in complex solutions parts or solutions in special complex detergent. detergent. Rinse with well distilled water water and dry at maximum 60°C in a dryer well with distilled water, and with water, well allow distilled to dry well with water well and distilled dry at maximum 60°C in a dryer 5% sodium Moisten a hypochlorite. clean tissue with water tissue and distilled wipe the well. surface as Proceed described above with Lubricate piston if neccessary using the piston Lubricate if neccessary that comes with that comes the pipette. compartment. Lubricate piston if neccessary using the compartment. grease piston Lubricate if neccessary Open pipette, parts rinse contaminated *(as *(as described in 3.) Open pipette and contaminated place torer's instructions. torer's Otherwise instructions. open Decontamination pipette, pipette, wipe with 90% ethanol, followed by by 2 M sodium and acetate again 90% ethanol.

water if distilled high-concentration water if high-concentration alkalis are alkalis water if high-concentration positive displacement systems should systems positive displacement be used Filtertips Filtertips should be used. 1. is different than Density that of water. It is advisable to occasionally rinse to It occasionally is advisable

the pipette and allow the liquid should should be positive used. Alternatively, the lower part of the pipette with lower part of the pipette with distilled acids are acids pipetted Using frequently.

Filtertips Filtertips is also recommended. rinse the to It occasionally is advisable 2. Pipetting should be carried out rapidly, positive displacement systems. positive displacement avoid Filtertips should contamination, To

the pipette. To avoid Filtertips or contamination, To

pipetted frequently. Using pipetted Filtertips Using is frequently. also recommended. avoid Filtertips contamination, To displacement systems can systems be used. displacement Therefore, it is necessary Therefore, to it adjust is necessary be used. An alternative would be to use be used. An alternative would be to use positive displacement systems. positive displacement distilled water. water. distilled due to the high vapor and pressure

Thermo guarantee sterility, To

3. After pipetting is finished, open changes in the changes wetting pressure. Pipettes Pipettes are with calibrated To avoid Filtertips should contamination, To to evaporate. Special features

Organic Acids infectious infectious Alkalis Proteins and buffers Nucleic Nucleic acids Potentially liquids solvents solutions solutions solutions Cell cultures Cell Liquid Handling Aqueous Radioactive

General Guidelines for Decontaminating Pipettes Pipettes Guidelines for General Decontaminating 16

Thermo Scientific Pipetting Guide General Guidelines for Decontaminating Pipettes Chemical Compatibility of Plastics

These are general guidelines, not performance guarantees. Factors such as concentration, temperature and length of exposure can affect performance. Pipetting Pipetting Guide Chemical Class Polypro- Polyethylene Polyvinylidene Silicone Polystyrene pylene (PP) (HD-PE) fluoride (PVDF) (PS) Finntip Plungers Tip cones BioMate Microtiter BioMate (Finntip Stepper, (BioControl, (adaptor) microplates of Plastics Compatibility Chemical (nose cone) Finntip PDP) Finnpipette Focus, Digital, Novus ) Acid, mineral Boric acid Good Good Good Good Good Chlorosulphuric acid Poor Fair Fair - - Hydrogen chloride 20% Good Good Good Fair Good Hydrogen chloride 25% Good Good Good Fair Fair Thermo Scientific Thermo Scientific Hydrogen fluoride 25% Good Good Good Poor - Nitric acid 70% Fair Good Good Poor Poor Perchloric acid Fair Good Good Poor Good Phosphoric acid 1% Good Good Good - Good Phosphoric acid 10% Good Good Good - Good Sulphuric acid 50% Good Good Good Poor Good Sulphuric acid 98% Poor Good Good Poor Poor Acid, organic Acetic anhydride Fair Good Poor Poor Poor Formic acid concentrate Good Good Good Fair Fair Lactic acid Good Good Good Good Good Maleic acid Good Good Good - Good Palmitic acid Good Good Good Poor Good Salicylic acid Good Good Good - Good Tannic acid Good Good Good Fair Good Alcohol Allyl alcohol Good Good Good - Good Amyl alcohol Good Good Good Poor Good Ethanol Good Good Good Fair Good Ethylene glycol 60% Good Good Good Good Good Ethylene glycol 100% Good Good Good Good Good Furfuryl alcohol Good Good Good Good Poor Glycerol Good Good Good Good Good Isobutanol Good Good Good Good Good Methanol Good Good Good Good Poor Aldehyde Acetaldehyde Good Good Good Fair Poor Formaldehyde 37% Good Good Good Fair Good Aliphatic hydrocarbon Heptane Fair - Good Poor Poor Hexane Fair Poor Good Poor Poor Amide Dimethylformamide Good Good Poor Fair Poor Acrylamide Good Good Good - Good Amine Aniline Good Good Good Fair Poor Pyridine Good Good Good Poor Poor Triethanolamine Good Good Good - Good Aromatic hydrocarbon Benzene Poor Fair Good Poor Poor Toluene Poor Fair Good Poor Poor Base Aluminum hydroxide Good Good Good - Good Ammonia concentrate Good Good Poor - Good Calcium hydroxide Good Good Fair Good Good Potassium hydroxide 10% Good Good Poor Fair Good Sodium hydroxide 10% Good Good Poor Fair Good Good = resistant, no effect Fair = limited resistance, only for short exposure Poor = not resistant - = no data available

17 ChemicalCompatibility Plasticsof Guide PipettingScientificThermo Chemical Compatibility of Plastics

Chemical Class Polypro- Polyethylene Polyvinylidene Silicone Polystyrene pylene (PP) (HD-PE) fluoride (PVDF) (PS) Finntip Plungers Tip cones BioMate Microtiter BioMate (Finntip Stepper, (BioControl, (adaptor) microplates (nose cone) Finntip PDP) Finnpipette Focus, Digital SCP, MCP) Ester Dibutyl phthalate Fair Good Good - Poor Ether Diethyl ether Fair Good Poor Poor Poor Polyalkylene glycol Good Good Good - - Polyethylene glycol Good Good Good - - Polyethylene sulfide Good Good Good - - Propylene oxide Good Good Good - Poor Halogenated hydrocarbon Bromochloromethane Poor Poor Poor Poor Poor Carbon tetrachloride Fair Poor Good Poor Poor 2-Chloroethanol Good Good Good - - Chlorobenzene Poor Fair Good Poor Poor Chloroform Fair Fair Good Poor Poor Dichloroethane Fair Fair Good - Poor Heterocyclic compounds Tetrahydrofuran Fair - Fair Poor - Ketone Acetone Fair Good Poor Poor Poor 2-Butanone - - Poor - - Methyl ethyl ketone Good Good Poor Poor Poor Phenol Phenol Fair Good Good Poor Poor Salt, inorganic Aluminum chloride Good Good Good Fair Good Aluminum fluoride Good Good Good Good Good Ammonium carbonate Good Good Good Fair Fair Barium chloride Good Good Good Good Good Calcium chloride Good Good Good Good Good Calcium sulphate Good Good Good - Good Copper (II) chloride 5% Good Good Good Good Good Iron (II) chloride Good Good Good Fair Good Iron (III) nitrate Good Good Good - - Iron (III) sulphate Good Good Good Fair Good Lithium bromide Good Good Good - - Magnesium chloride Good Good Good Good Good Magnesium (I) nitrate Good Good Good - - Mercury (II) chloride Good Good Good - Good Nickel nitrate Good Good Good Good - Potassium carbonate Good Good Poor - Good Potassium chlorate Good Good Poor Fair - Silver nitrate Good Good Good Good Fair Sodium carbonate Good Good Good Good Good Sodium fluoride Good Good Good - - Sodium hypochlorite 5% Fair Good Fair Fair Good Tin (II) chloride Good Good Good Fair - Tin (IV) chloride Good Good Good Fair - Zinc chloride Good Good Good Good Good Zinc sulphate Good Good Good Good - Miscellaneous Culture Media Good Good Good Good Good DEPC Good - Good Good Good DMSO Good Good Poor - Good Ficoll-Hypaque Good Good Good Good Good Sera Good Good Good Good Good Urea Good Good Good Fair Good

Good = resistant, no effect Fair = limited resistance, only for short exposure Poor = not resistant - = no data available

18 Customer support Customer support Customer

You can enter your questions concerning Finnpipettes and Finntips to our Customer Support Desk. To do so, Pipetting Guide please go to our web site www.thermo.com/finnpipette -> Service & Support -> Customer Support Desk. You will have to register to be able to send your inquiries. Thermo Scientific Thermo Scientific

Troubleshooting

Defect Possible reason Solution Leakage Tip incorrectly attached Attach firmly Foreign particles between Clean tip cones attach tip and tip cone new tips Foreign particles between Clean and grease O-ring the piston, the O-ring and and cylinder the cylinder Insufficient amount of grease Grease accordingly on cylinder and O-ring O-ring damaged Change the O-ring

Inaccurate dispensing Incorrect operation Follow instructions carefully Tip incorrectly attached Attach firmly

Calibration altered: e.g. Recalibrate according to instructions caused by misuse Inaccurate dispensing Unsuitable calibration Recalibrate with the with certain liquids High viscosity liquids may liquids in question require recalibration

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