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ถกูต้อง“Guide to Pipetting”694 ซ.รชัดานิเวศน์ 24 ถ.ประชาราษฎรบ์ำาเพญ็ เขตห้วยขวาง
กรุงเทพฯ 10321Tel. 0-2274-2456 (automatic 9 lines) Fax. 0-
2274-2443 Email. [email protected]
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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