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บรษัิทหรกิลุ ซายเอนซ ์จำากัดเทคนิคการใชง้านปิเปตอยา่ง

ถกูต้อง“Guide to Pipetting”694 ซ.รชัดานิเวศน์ 24 ถ.ประชาราษฎรบ์ำาเพญ็ เขตห้วยขวาง

กรุงเทพฯ 10321Tel. 0-2274-2456 (automatic 9 lines) Fax. 0-

2274-2443 Email. pipetman@harikul.com

Choosing the right pipette

Air – displacement

Positive – displacement

Two families for Gilson pipettesAir-displacement Positive-displacement

Operating principles

• Air - displacement • Positive - displacement

Select the best pipetting system/sample

Air-displacement Positive-displacement

Air-displacement

Forward Mode

Reverse Mode

Rest position

First stopSecond stop or purge

Air-displacement pipettesforward pipetting mode

Rest position

First stopSecond stop or purge

Air-displacement pipettes reverse pipetting mode

Positive-displacement

Capillary Pistons (CP)

Positive-displacement

1/ Preparation

2/ Aspiration 3/ Distribution 4/ Ejection

Rest position

First stopEjection

How to handle a pipette

Pipetting cycle

1.Fit a tip2.Set the volume3.Pre-rinse4.Aspirate5.Dispense6.Eject7.Hold the pipette

Fit a tip

multichannelsSingle channels

Set the volumeSet the volume

- using the thumbwheel or the push-button

Electronic display avoids parallax error

For Pipetman Classic use the horizontal positionand place your eye in front of the volumeter

Pre-rinse

Aspirate with the tip and dispense back

• Why?– Wettability of the tip Greater uniformity Greater accuracy and precision

• When?– After changing the tip– After increasing the volume

Aspirate

Dispense

Eject

Hold the pipette

Improve your working day

Get organized J Necessary objects within easy arm's reach Pipettes within easy reach, on a stand Most frequently used objects in front of you Waste receptacles no higher than the end of your pipette

Posture J Chair or stool so that elbow on the bench Hands below shoulder height Reduce the height of applications such as gel loading Arms close to the body

PostureGood J

Seated posture:Lower back supported by chairUpper back and neck uprightUpper arm verticalElbow bent at 90°Wrist in the same plane as the forearmStanding posture:Lower back and trunk uprightUpper back and neck uprightUpper arm verticalElbow bent at 90°Forearm parallel to the floorWrist in the same plane as the forearm

Wrist posture:Forearm parallel to the floorWrist and forearm in the same plane

L Bad

Seated posture:Shoulders elevatedUpper arm elevatedElbow extendedWrist in deviation

Standing posture:Upper back and neck stoppedLower back and trunk stoppedElbow flexed

Wrist posture:Upper arm flexedElbow extendedWrist deviated downward

Regular maintenance

Leak testFrom 1000 µL to 10 mLFrom 0.2 µL up to 200 µL

Step 1 Step 2 Step 1

CleaningExternal

Remove the tip ejectorWipe with soap solutionWipe with distilled water

Tip ejector

Connecting nut

Seal & O-ring

Piston assembly

InternalDisassemble the lower partClean then rinse the tip holder,

piston assembly and sealsLeave to air dry and reassemble

Disassemble & Reassemble

Autoclaving

lower part is autoclavable connecting nut; tip ejector; shaft/tip holder Moist heat/121°C/20 minutes/1 bar

Maintenance frequency

Frequency * Control Action Who

Daily Preventive maintenance Leak test (See the "Two-minute inspection") End-usersWeekly Preventive maintenance Accuracy check End-users

Every 3 months Preventive maintenance

Cleaning & Diagnosis: - Visual inspection - Check functions

End-users

Annually

Replacement spare parts of first level (seals/ O-rings, tip holder…) /Adjustment / Calibration (3x4; 3x10 measurements)

Service Center & End-users

Replacement spare parts of second level (Piston, volumeter, operat ing rod) /Adjustment / Calibration (3x4; 3x10 measurements)

Gilson Authorized Service Center

Complete maintenance

Performances check

Specifications

Performances

Accurate and precise

Systematic error

Accuratebut not precise

Precisebut not accurate

Random error

Errors

Random errorTipsTechnique Piston

Systematic errorZ factor not adequateNeeds adjustment

ENVIRONMENT

EQUIPMENT

TECHNIQUE

TIPS

Parameters

AIR TEMPERATUREWATER TEMPERATURE

AIR CONDITIONNING

AIR PRESSURE

DENSITY

DRAUGHTSSTABILITY

DIRECT SUNHUMIDITY

WATERAir buoyancy

Repeatability

MARBLE TABLE

Weighing vessel

BALANCESPECIFICATIONS

MAINTENANCE

READIBILITY

CLASS

LINEARITY

CONTROLSRECORDER SIZE

ShapeLID

PERIODICAL CHECKSSTANDARD MASS

Drift

VibrationsSPEED

OPERATING MODE

QUALIFICATION

CONSISTENCY

EVAPORATION

IMMERSION DEPTH

PRE RINSING

StabilizationRegularity

Smooth and regularWETTABILITY

Perfectly formed

AIR-TIGHT FITTRACEABILITY

Fine point finishClean

Sources of errorInfluencing parameter Effect Solution

Temperature (pipette, room, liquid)

up to 0.3% Same temperature

Uneven rythm and timing up to 1.5% Even technique

Failure to prewet tip up to 2.0% Pre-wetting of pipette tip

Immersion depth and handling angle

up to 1.0%Vertical position, user

information

Wipe pipette tip on the wall up to 3.0% Wiping distance ~ 8 to 10mm

Tips Leaky/poorly seated 0.5% to 50% Use recommended pipette tips

EquipmentBalance resolution and

linearitybalance linearity

check balance with standard mass and choose your

balance according to the

Technique

Environment

Excess plastic

Imperfect point aperture

Rough surface

Rough surface

Pipette Tips

Identification: mold and cavity

Level marks

Sealing ring for airtight fit

Translucent, no dyes

Fully autoclavable, even the filter

Fine pointand flexibility

L

JPerfect point

Non Gilson Tips

Gilson Diamond Tips

calibration method

• Test Volume: minimum and Maximum

• Test Volume: minimum Medium and Maximum

Harikul Calibration Laboratory

Gravimetry method

Equipment : Balances requirement

Volume (ul) HARIKUL ISO 8655

0.1 ul.≤V 1 ul. 0.0001 mg -1 ul ≤V≤10 ul 0.0001 mg 0.001 mg.

10 ul V≤20 ul 0.0001 mg 0.01 mg.20 ul V≤ 100 ul 0.001 mg 0.01 mg.

100 ul V≤ 1000 ul 0.01 mg. 0.1 mg.1 ml V≤ 10 ml 0.01 mg. 0.1 mg.

10 ml V≤ 200 ml 0.01 mg. 1 mg.

Environmental factors to be respected

101kPa101kPa

22.5 ±7.5°C20.0 + 2°C

Above 50%40% to 60%

ISO 8655Harikul Calibration Laboratory

2320

Do not forget: Conversion factor Evaporation loss

Questions / Comments

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