EVOLVE Manual Pipettes Operating instructions
900-00010-00_V00
Declaration of conformity
INTEGRA Biosciences Corp. –
Hudson, NH, USA
declares on its own responsibility that the devices
comply with:
Hudson, New Hampshire, USA - March 15, 2016
Description Models
EVOLVE Manual
Pipettes
3011, 3012, 3013, 3015, 3016, 3018, 3019, 3022, 3024, 3025,
3026, 3032, 3034, 3035, 3036
EU Directives (DoW: Date of Withdrawal) Before DoW DoW After DoW
Restriction of Hazardous Substances 2011/65/EU
EU Regulations
Registration, Evaluation, Authorisation and Restriction of Chemicals
(REACH)
1907/2006
Piston-operated volumetric apparatus - Part 2: Piston pipettes ISO 8655-2
George Kalmakis
VP Operations
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Table of Contents
Chapter 1 Introduction
1.1 Symbols used...................................................................................5
1.2 Intended use.....................................................................................5
1.3 Safety notes .....................................................................................6
Chapter 2 Description of the device
2.1 Scope of delivery..............................................................................7
2.2 Overview of the EVOLVE Manual Pipettes ......................................7
Chapter 3 Installation
3.1 Optional color coded plunger cap.....................................................8
3.2 Changing the Plunger Spring ...........................................................8
Chapter 4 Operation
4.1 Setting the volume..........................................................................10
4.2 Attaching and removing GripTip pipette tips .................................. 10
4.3 Pipetting .........................................................................................11
4.3.1 Aspirating and Dispensing ..................................................11
4.3.2 Reverse mode.....................................................................12
4.3.3 Proper pipetting techniques ................................................13
4.4 Troubleshooting..............................................................................14
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Chapter 5 Maintenance
5.1 Cleaning .........................................................................................15
5.2 Single channel lower end disassembly ..........................................15
5.2.1 Disassembly........................................................................15
5.2.2 Reassembly ........................................................................18
5.3 Sterilization.....................................................................................20
5.3.1 Autoclaving the disassembled components........................20
5.4 Servicing.........................................................................................21
5.4.1 Shipping to INTEGRA Biosciences.....................................21
5.4.2 Changing O-rings of tip fittings............................................21
5.4.3 Lubrication ..........................................................................22
5.5 Calibration ......................................................................................23
5.5.1 Materials .............................................................................23
5.5.2 Definitions ...........................................................................23
5.5.3 Test conditions and environment ........................................24
5.5.4 Pre-wetting of tips ...............................................................24
5.5.5 Leak test .............................................................................25
5.5.6 Obtaining the actual volume ...............................................26
5.5.7 Calculation of accuracy and precision ................................27
5.5.8 Adjusting EVOLVE Manual Pipettes...................................28
5.6 Equipment disposal ........................................................................29
Chapter 6 Technical Data
6.1 Environmental conditions ...............................................................30
6.2 Pipette specifications......................................................................30
6.3 Z Correction Factors.......................................................................32
Chapter 7 Accessories and Consumables
7.1 Accessories ....................................................................................33
7.2 Consumables .................................................................................33
Imprint................................................................................................... 36
EVOLVE Manual Pipettes – Operating instructions V00 Introduction
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1 Introduction
These operating instructions contain all the information required for installation, operation
and maintenance of the EVOLVE Manual Pipettes. This chapter informs about the
symbols used in these operating instructions, the intended use of the EVOLVE Manual
Pipettes and the general safety instructions.
1.1 Symbols used
The operating instructions specifically advise of residual risks with the following symbols:
W
ARNING
This safety symbol warns against hazards that could result in injury. It also
indicates hazards for machinery, materials and the environment.
It is essential that you follow the corresponding precautions.
C
AUTION
This symbol cautions against potential material damage. Follow the
instructions.
N
OTE
This symbol identifies important notes regarding the correct operation of the
device and labor-saving features.
1.2 Intended use
EVOLVE Manual Pipettes are intended to be used in a laboratory for aspirating and
dispensing liquids in the volume range of 0.2–5000 µl using GripTip pipette tips.
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1.3 Safety notes
EVOLVE Manual Pipettes comply to the recognized safety regulations and is safe to
operate. EVOLVE Manual Pipettes can only be operated when in perfect condition and
while observing these operating instructions.
The device may be associated with residual risks if it is used or operated improperly by
untrained personnel. Any person operating the EVOLVE Manual Pipettes must have read
and understood these operating instructions, and particularly, the safety notes, or must
have been instructed by supervisors so that safe operation of the device is guaranteed.
Repairs may only be performed by INTEGRA Biosciences AG or by an authorized aftersales service member.
Parts may be replaced with original INTEGRA Biosciences parts only.
Regardless of the listed safety notes, additional applicable regulations and guidelines of
trade associations, health authorities, trade supervisory offices, etc. must be observed.
Please visit our website www.integra-biosciences.com
on a regular basis for up to date
information regarding REACH classified chemicals contained in our products.
W
ARNING
When handling dangerous substances, comply with the material safety data
sheet (MSDS) and with all safety guidelines such as the use of protective
clothing and safety goggles.
CAUTION
Do not immerse the EVOLVE Manual Pipettes in liquid. The fluid can damage
internal parts. Avoid pipetting of liquids whose vapors could attack the
materials PA (polyamide), POM (polyoxymethylene), FPM (fluor-rubber),
NBR (nitrile-rubber), CR (chloroprene), silicone. Corrosive vapors could also
damage metallic parts inside the device.
N
OTE
Prolonged exposure of the EVOLVE Manual Pipettes to UV-light can cause
discoloration and/or yellowing of the pipette housing. However, this will not
affect the performance of the device in any way.
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2 Description of the device
2.1 Scope of delivery
• EVOLVE Manual Pipette
• Multipurpose Tool
• Replacement Plunger Cap
• Quick Start Guide
• Replacement O-rings (for 200 μl, 1000 μl and 5000 μl models)
2.2 Overview of the EVOLVE Manual Pipettes
1 Plunger Cap
2 Plunger, to pipette
3 Tip Ejector
4 Finger Hook
5 Adjustment Dials, to set volume
6 Volume Window
The color of the volume indicator label matches
GripTip box insert
7 Ejector Sleeve
8 Tip Fitting
1
2
4
6
3
5
7
8
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3 Installation
3.1 Optional color coded plunger cap
If desired, replace black plunger cap with optional color coded plunger cap.
3.2 Changing the Plunger Spring
The plunger spring installed inside the pipette is the light spring option. You can exchange
with a heavier plunger spring to achieve more control of the plunger movement.
1) Unscrew the Cylinder Assembly (a) and
separate the Piston Assembly (b, models 2–
200 µl) or Piston (model 1000 µl) from the upper
part of the pipette (c). It is held in place by a small
magnet.
N
OTE
5000 µl piston and cylinder assembly is
removed as one piece.
2) Set the pipette to the maximum setting. The
maximum setting is higher than the maximum
volume.
3) Put the two teeth of the Multipurpose Tool (d)
vertically into the slots of the black opening inside
at the bottom of the pipette‘s body (c). Rotate
counterclockwise until the plunger spring
assembly (e) is released.
a
b
c
d
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CAUTION
Validate or calibrate your pipette after reassembly.
4) To install the optional spring assembly, screw the
plunger spring assembly (e) clockwise into the
upper part of the pipette using the Multipurpose
Tool.
5) Re-attach the Piston assembly/Piston. Screw
clockwise to attach the Cylinder Assembly (a) to
the body of the pipette.
e
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4 Operation
4.1 Setting the volume
4.2 Attaching and removing GripTip pipette tips
CAUTION
T o ensure optimal performance of your EVOL VE Manual Pipettes always use
GripTips of correct size which have been designated for the use with the
EVOLVE Manual Pipettes, see “7.2
Consumables” on page 33.
The unique GripTip system of the EVOLVE Manual Pipettes reduces attachment and
ejection forces, ensures a perfect fitting to prevent the tips from falling off and to provide
a perfect seal. On a multichannel pipette all tips sit on the same height.
Attach the tips:
When loading tip(s), press the pipette into the appropriate GripTip(s) until you hear and
feel a click indicating that a seal has been achieved. Once you feel the click, stop applying
pressure. When loading GripTip pipette tips using a multichannel pipette, slowly rock the
pipette from one side to the other side to ensure the proper seal is achieved.
Discard your used tips:
If liquid is in the tips, empty them by pressing down the Plunger (2
) to the second stop
(see section 4.3
). Tips are easily removed by pressing the Tip Ejector (3).
1) To set the desired pipetting volume press down the Plunger (a)
and rotate slightly clockwise to unlock the volume dials.
2) Spin the Adjustment Dials (b) until the desired volume is
shown in the Volume Window.
3) Slightly rotate and slowly release the Plunger (a) to lock the
volume.
a
b
A rim inside the GripTips snaps over the multi-lobes and ensures
firm attachment of the tips.
A shoulder provides a positive stop to prevent over tightening of the
tips. Without hammering, the tip is either on or off but nothing in
between.
O-ring provides forgiving and robust seal surface for the pipette tip.
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4.3 Pipetting
The Plunger can be pushed down to two different stops:
For detailed recommendations for pipetting see also “4.3.3
Proper pipetting techniques”
on page 13.
4.3.1 Aspirating and Dispensing
To aspirate and dispense liquid perform the following steps:
1) Top position: Varies depending on volume setting.
2) First stop: Fixed calibrated position for aspirating and
dispensing.
3) Second Stop: To blowout the residual liquid.
Aspirate
1) With GripTip(s) attached, push down the Plunger (2
) to the first
stop.
2) Place GripTips in liquid. Hold the pipette vertically or in an angle
of max. 20° throughout the entire pipetting process and immerse
the GripTip(s) just below the surface of the liquid (2–3 mm).
3) Slowly return Plunger to the top position. For higher volumes
(>200 μl), follow the liquid level by moving the pipette down.
4) Remove the GripTip(s) from liquid.
Dispense
5) To dispense the liquid, push the Plunger (2
) down to the first
stop.
6) If there is any residual liquid in tip, a blowout
should be
performed by pressing the Plunger to the second stop.
7) Remove the GripTips from vessel and release Plunger.
1
2
3
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4.3.2 Reverse mode
Viscous samples should be aspirated and dispensed at slower speeds and in reverse
pipetting mode:
Aspirate
1) Fully push down the Plunger (2
) to the second stop.
2) Immerse the GripTip(s) in the liquid, let the Plunger slowly
release to the upper position and wait shortly until the liquid is
fully aspirated.
Dispense
3) To dispense, press the Plunger (2
) slowly down to the first stop
and wait a few seconds to dispense the liquid.
4) Remove the GripTip(s) from the target vessel.
5) To expel the excess liquid press the Plunger to the second
stop.
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4.3.3 Proper pipetting techniques
INTEGRA Biosciences recommends the following techniques for enhancing pipetting
results. These techniques are consistent with ISO standard 8655-2.
• It is best to immerse the GripTips just below the surface of the liquid (2–3 mm) to allow
the desired volume to be aspirated. If the tip is immersed too deep, the risk of carryover is increased due to liquid drops clinging to the outer surface of the tip.
• Press and release the plunger with consistent speed and smoothness.
• Always prewet GripTips. After loading tips onto your pipette, aspirate and dispense the
full volume 2-3 times to coat the inside of pipet tips. Pre-wetting ensures that the liquid
and air inside the tips are at equal temperature and the dead air space is humidified.
Neglecting to perform a pre-wet can result in a smaller delivery volume in the first few
dispenses.
• EVOLVE Manual Pipettes are air displacement pipettes. To properly dispense liquids,
ensure that the pipette tip is at a 0–20° angle. After each dispense, you must touch
GripTips against the vessel wall or liquid. This process is referred to as “touching off”
or “tip touch” and prevents liquid from clinging to the pipette tips.
• Air displacement pipettes show the best performance between 35% and 100 % of the
nominal volume.
• Viscous samples should be aspirated and dispensed at slowest speeds to ensure
accurate pipetting. In addition, pipetting should be performed in a reverse mode.
• For pipetting liquids with high vapor pressures (such as methanol or ethanol), use relatively fast pipetting speeds and avoid prolonged pauses after aspiration.
• Calibrate based on fluid type. EVOLVE Manual Pipettes are tested and calibrated at
the factory for use with distilled water at room temperature. It may be necessary to recalibrate your EVOLVE Manual Pipettes if the liquid to be used has different physical
properties (specific gravity and vapor pressure) than water.
W
ARNING
Avoid pipetting for extended periods. To minimize the risk of repetitive strain
injury, include pauses of several minutes.
Use filtered GripTips to pipette corrosive or biohazardous liquids. The filter
prevents vapors and aerosols from corroding or contaminating the cylinder
assembly.
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4.4 Troubleshooting
Problem Probable cause Remedy
Leaking. • Tip incorrectly attached.
• Foreign particles between tip
and tip fitting.
• Damaged red O-ring.
• Attach a new tip.
• Clean tip fitting. Attach new tips.
• Change the O-ring, see 5.4.2
.
• If leak persists, contact service.
Dispense results
are inaccurate.
• Unsuitable calibration for
liquid class.
• Improper pipetting
techniques.
• Recalibrate with the liquids in
question.
• Adjust aspiration and dispense
speed depending on liquid:
- High viscosity liquids may
require calibration.
- High vapor pressure liquids
may require pre-wetting.
- Refer to proper pipetting
techniques section 4.3.3
Not dispensing/
aspirating.
• Piston stuck or not
connected.
• Internal seal is damaged.
• Contact service.
Droplets on the
tips.
• Temperature of liquid differs
from that of air inside the tips.
• Liquid of low viscosity and
high vapor pressure.
• Touch off was not performed.
• Pre-wet tips up to 3 times.
• Increase dispensing speed.
• Perform a touch-off.
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5 Maintenance
5.1 Cleaning
The materials used on the exterior of the EVOLVE Manual Pipettes support regular
cleaning intervals. Clean the external components with a lint-free cloth lightly soaked with
mild soap solution in distilled water or with a 70 % dilution of Isopropyl or Ethanol. Never
use acetone or other solvents.
W
ARNING
Do not immerse the entire pipette into a cleaning solution or spray cleani ng
solution directly onto the exterior body of the pipette.
If the pipette is exposed to corrosive vapors, e.g. from concentrated acids, or if liquid
enters the internals, disassemble the lower part of the pipette and clean the internal parts
with distilled water or with a 70 % dilution of Isopropyl or Ethanol. Follow the instructions
below for disassembly, also necessary for trouble shooting and autoclaving.
5.2 Single channel lower end disassembly
5.2.1 Disassembly
Disassemble the lower part of the single channel pipette as follows:
Models of all sizes:
1) Unscrew counterclockwise to remove the
Cylinder Assembly (a) from the pipette.
2) Slide the Cylinder Assembly (a) down from the
pipette body. This exposes the Piston Assembly
(b, models 2–200 µl) or Piston with mounted
Cup Seal (m, model 1000 µl).
a
b
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Models 2–20 µl:
3) Slide the following components from the Piston (h):
•
O-Ring
(black, g)
and Seal
(white, f)
Assembly
• Lower Seal Retainer (white, e)
• Seal Retainer Spring (d)
• Seal Retainer (white, c)
Set these components aside or place them in an
autoclave pouch.
Models 100 µl and 200 µl:
3) Slide the following components from the Piston (h):
•
O-Ring
(black, g)
and Seal
(white, f)
Assembly
• Seal Retainer Spring (d)
• Seal Retainer (black or white, c)
Set these components aside or place them in an
autoclave pouch.
100 µl 200 µl
Models 2–1000 µl:
4) Separate the Piston (h) from the upper part of the
pipette. The Piston is held in place by a small
magnet.
f
c
d
h
e
g
f
d
c
h
g
f
d
c
h
g
h
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Models of all sizes:
6) Remove the Ejector Spring (j).
7) Set all components of the lower part aside or place them in an autoclave pouch.
Cylinder Assemblies (a): Models 2–1000 µl:
5) With the Cylinder Assembly (a) in hand, turn
counterclockwise to remove the black Snap Ring
(i). Pullout the black Cylinder (k) from the white
Ejector Sleeve (l).
Model 5000 µl:
5) Put your fingernails into the gap of the white
Ejector Sleeve (l) and pull off the Ejector Sleeve
Cap (n). With the black Cylinder (k) in hand
unscrew the upper part. Remove the Piston (h)
with mounted Cup Seal (m).
2–1000 µl
5000 µl
k
j
i
l
h
l
n
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5.2.2 Reassembly
Before reassembling the pipette, check each component for lint or dust particles. It is
recommended to replace the O-Ring (black, g) and Seal (white, f) or Cup Seal (m).
1) Lightly lubricate the Piston, O-Rings and Seals (see “5.4.3
Lubrication” on page 22).
Models 2 µl –1000 µl:
2) Position the Piston (h) at the small magnet to connect the Piston to the upper part of
the pipette.
Models 2 µl – 200 µl:
3) Slide the Seal Retainer (c) onto the Piston and up into the pipette. The end of the Seal
Retainer that has an extended edge (lip) faces the upper part of the pipette.
4) Slide the Seal Retainer Spring (d) onto the Piston. It rests against the Seal Retainer.
Slide the Lower Seal Retainer (e, 2, 10, 20 µl only) onto the Piston.
5) Slide the lightly lubricated
O-Ring
(black, g)
and Seal
(white, f)
Assembly onto the
Piston. The white Seal is closest to the Seal Retainer Spring. Be sure the black O-Ring
is securely slided into the white Seal.
Piston Assemblies (models 2–200 µl) and Pistons with Cup Seal (1000 µl, 5000 µl):
c: Seal
Retainer
d: Seal
Retainer
Spring
e: Lower Seal
Retainer
f: O-Ring
g: Seal
h: Piston
m: Cup Seal
2, 10, 20 µl 100, 200 µl 1000 µl 5000 µl
g
c
d
h
e
f
f
d
c
h
g
h
m
m
h
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Models of all sizes:
6) Slide the Ejector Spring (j) onto the top of the black Cylinder (k).
Models of all sizes:
10) Screw clockwise to attach the Cylinder Assembly (a) to the body of the pipette.
Perform a leak test (see “5.5.5
Leak test” on page 25) and validate pipetting volumes after
reassembly.
Cylinder Assemblies (a): Models 2 µl – 1000 µl:
2–1000 µl
5000 µl
7) Slide the Cylinder (k) into the white Ejector Sleeve (l).
8) Position the black Snap Ring (i) over the Cylinder (k) at
the top of the Ejector Sleeve (l). Turn the sleeve
clockwise until the Snap Ring snaps into place and
secures the Cylinder in the Cylinder Assembly.
9) Slide the Cylinder Assembly (a) over the Piston (h).
Model 5000 µl:
7) Push the Cup Seal (m) of the Piston (h) into the top of
Cylinder‘s lower part (k) and screw on the upper part of
Cylinder (k).
8) Ensure that the four noses (o) of the Ejector Sleeve Cap
(n) point to the openings (p) of the Ejector Sleeve (l) and
re-attach the cap.
k
j
i
l
n
h
o
p
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5.3 Sterilization
If the surface of the EVOLVE Manual Pipettes have been in contact with biohazardous
material, they must be decontaminated in accordance to good laboratory practice. Wipe
the clean surface with a lint-free cloth, lightly soaked e. g. with the following disinfectants:
• Ethanol 70 %
• Microcide SQ 1:64
• Glutaraldehyde solution 4%
• Virkon solution 1-3%
Follow the instructions provided with the disinfectants.
It is not recommended to autoclave the EVOLVE Manual Pipettes. If autoclaving is
required, only the lower assembly of the EVOLVE single channel pipettes can be
autoclaved.
W
ARNING
Service is required after autoclaving the EVOLVE Manual Pipettes!
Do not autoclave the entire unit. Multichannel pipettes can NOT be
autoclaved, they might be damaged!
As-found calibration (measurement report, indicating “before” data) is not
possible after autoclaving!
5.3.1 Autoclaving the disassembled components
Place the disassembled components (see 5.2.1) into steam autoclave in an autoclave
pouch:
You may autoclave the components at 121°C, 1 bar overpressure for 20 minutes.
C
AUTION
Service is required after autoclaving the EVOLVE Manual Pipettes!
Single channel 2–1000 µl: Single channel 5000 µl:
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5.4 Servicing
5.4.1 Shipping to INTEGRA Biosciences
For any service or repairs, please contact your local service technician.
WARNING
If working with infectious materials, e. g. human pathogens, the EVOLVE
Manual Pipettes need to be decontaminated before sending them to service
and the declaration on the absence of health hazards must be signed. T his
is necessary to protect service personnel.
5.4.2 Changing O-rings of tip fittings
200 µl, 1000 µl and 5000 µl EVOLVE Manual Pipettes feature tip fittings with red O-rings.
This O-ring is used to seal against the inside wall of GripTips and provides a robust seal.
O-rings are made of durable silicone. If necessary, e.g. in case of a leakage due to
damaged O-ring, you can replace these O-rings. A set of spare O-rings can be ordered
separately, see “7.2
Consumables” on page 33.
W
ARNING
Avoid mechanical damage of the tip fittings.
For 200 µl or 1000 µl pipettes, slide the Multipurpose Tool
sidewards onto the tip fitting until the O-ring (a) builds a loop.
Remove the O-ring with plastic tweezers.
For 5000 µl pipettes, simply use plastic tweezers to remove the Oring.
Slide a new O-ring over the tip fitting (b).
a
a
b
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5.4.3 Lubrication
The internal seals and O-rings are subject to wear. An undamaged thin lubricant film is
important to keep the seals tight. The lubricant recommended depends on the EVOLVE
pipette size, see “7.2 Consumables” on page 33:
Single channel 2 µl– 200 µl models:
Use Fluorocarbon Gel (#100-00136-50) to lightly lubricate the
piston (a) without components (c, d) slid onto. Put a drop of grease
onto your fingers, lubricate the black O-ring (b) and slide it over the
white seal (c).
Single channel 1000 µl and 5000 µl models:
Use the Super-O-Lube (#100-00135-50) to lightly lubricate the
outer ring of the Cup Seals (d, e). Do not lubricate the bottom of Cup
Seals.
b
c
a
d
e
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5.5 Calibration
The EVOLVE Manual Pipettes are factory tested and calibrated under environmentally
controlled conditions using a gravimetric procedure in accordance with ISO 8655
standards. For the accuracy and precision specifications, see “6.2 Pipette specifications”
on page 30.
For information regarding calibration service, please contact your local dealer.
5.5.1 Materials
• Precision balance with 0.01 mg readability
• ASTM Class 2 or OIML E2 test weights
• Evaporation trap for balance
• Weighing vessel (optimally the height-to-diameter ratio is at least 3:1)
• Measurement equipment for temperature, humidity and atmospheric pressure
• Distilled water (Grade 3 according to ISO 3696)
• New non-filtered GripTips matching the pipette volume (see section 7.2
)
• 1 beaker with distilled water
5.5.2 Definitions
Blow-out: Discharges any residual liquid from the pipette tip.
Blow-in: After the blow-out, the piston moves back into home position. This causes a
slight intake of air (or liquid if tips remain in the liquid).
Two step blow-out: To avoid an early blow-in, keep the plunger pressed after dispensing,
remove the tips from liquid and then release the plunger.
Pre-wet: The action of pre-coating the inside of the liquid contacting parts with a thin film
of the same liquid. Additionally, it equilibrates humidity of the air space inside tip and the
pipette.
Touch off: Touching the pipette tip against the surface liquid or side of well-plate to
release any sample liquids that might be on the end of the pipette tip.
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24 INTEGRA Biosciences AG
5.5.3 Test conditions and environment
Tests and calibrations should be performed in conditions and environment according to
ISO 8655-6 standard.
• Temperature needs to be between 18–25°C and remain constant (±0.5 °C) throughout
the calibration.
• Optimal relative humidity of the environment is >50% and around the dispensing position 80%.
• GripTips, pipettes and distilled water need to be in the calibration laboratory for at
least 2 hours prior to calibration to reach temperature equilibrium with the environment.
• The balance must be validated using reference weights before and after a measuring
series. A low and high volume weight should be used. E.g.:
100 g (Mettler Toledo, #11119250)
10 g (Mettler Toledo, #11119220)
1 g (Met tle r Toled o, #11119190)
10 m g (M ett ler Tol edo, #1111 913 0)
C
AUTION
Always use new, pre-wetted GripTips for leak test and calibration.
5.5.4 Pre-wetting of tips
Pre-wet new GripTips three times prior to starting tests and calibrations. This is required
every time a tip is changed.
1) Set the pipette to aspirate full volume.
2) Attach a new unused GripTip which correspond to the volume range of the pipette
directly from the rack without touching the tip by hand.
3) Use a separate waste container for pre-wet dispenses.
4) Push down the Plunger to the first stop. Aspirate by releasing with medium speed.
5) Dispense: Keep the Plunger pressed to the first stop until all liquid is expelled.
At the end of the third cycle push the Plunger to the second stop, remove the tip from the
liquid and then release the Plunger to perform a two-step blowout.
EVOLVE Manual Pipettes – Operating instructions V00 Maintenance
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5.5.5 Leak test
It is recommended to perform a leak test every 3 months or when errors occur.
1) Prewet tips as described above.
2) Set the EVOLVE Manual Pipettes to aspirate full volume.
3) Push down the Plunger to the first stop. Aspirate full volume by releasing with medium
speed and verify liquid level is not decreasing during 10 seconds while GripTips are
still in distilled water.
4) Remove the GripTips from the liquid and hold the pipette at a 20° angle. Wait for 20
seconds.
a) Observe whether liquid droplets are forming at the end of the GripTips.
b) On a multichannel pipette, verify liquid level is even across all channels.
5) Immerse GripTips approximately 2 mm into distilled water and push and release the
Plunger to mix six times.
a) Observe whether air bubbles are forming when dispensing.
b) On a multichannel pipette, check whether the levels stay approximately on the
same level across all channels.
6) At the end push the Plunger to the second stop and remove the tip from the liquid
before releasing.
Signs indicating a leak
1) During the mix cycle in the leak test, the liquid level of 1 channel is decreasing.
N
OTE
A decreasing liquid level at aspiration could be an indication of a slow leak.
Performing a retest at 10 mixes may help identify a slow leak.
2) Liquid is left in a tip after the last dispense during the leak test.
3) One or more channels show air bubbles during the mix cycle in the leak test.
4) Droplets are forming when holding the pipette in the air for 20 seconds, even if a prewet was performed.
5) The liquid levels are not equal on all channels after aspiration.
If the pipette is leaking, change O-rings (see 5.4.2
) and lubricate pistons (see 5.4.3) or
contact your service technician.
EVOLVE Manual Pipettes – Operating instructions V00 Maintenance
26 INTEGRA Biosciences AG
5.5.6 Obtaining the actual volume
INTEGRA certifies the pipettes at 10 %, 50% and 100 % of the nominal value. For each
volume 5 measurements are taken from all channels.
General
1) Always use new, unused GripTips, also when changing the test volume (e.g. from
200 µl to 20 µl).
2) Always pre-wet (section 5.5.4
) when using a new GripTip. For a low volume
measurement, first set the maximum volume for the pre-wet and then change to the
volume to be measured.
3) After dispensing, perform a touch off to make sure no liquid remains on the tip.
Gravimetric testing
1) Write down the ambient temperature and air pressure.
2) Set the pipette to the high test volume.
3) Perform a pre-wet, see 5.5.4
.
4) The first and second dispense to the balance should not be recorded. After each
dispense re-tare the balance.
5) Aspirate the Target volume of water keeping the pipette in a vertical to 20-degree
position while immersing the pipet tip 2–3 mm below the surface of the water. When
withdrawing the tip from the liquid, gently wipe the tip against the side wall of the vessel
to remove any liquid from the outside of the pipet tip.
6) Start the first measurement. Always pipet directly into the liquid of the weigh container
on the balance. At the end push the Plunger to the second stop and hold the plunger
pressed until the pipette is removed again from the weigh vessel to perform a 2-step
blowout. Record the weight from the balance.
7) After completing the 5 high volume measurements, continue with the mid and low test
measurements by repeating steps 3–6.
EVOLVE Manual Pipettes – Operating instructions V00 Maintenance
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5.5.7 Calculation of accuracy and precision
Accuracy
The ability of a pipette to aspirate or dispense the exact volume desired. It indicates the
proximity of measurement results to the true value. Accuracy is also know as systematic
error and as the name indicates, can be corrected/calibrated.
Precision, coefficient of variation (CV)
Precision indicates the repeatability or reproducibility of the measurement. It is also called
random error and is therefore an unpredictable error, which can not be corrected/
calibrated.
Conversion of the mass to volume
The values obtained by balance readings are in grams. These values need to be
converted to micro liters using the Z correction factor. It takes into account the water
density and air buoyancy during weighing at the corresponding test temperature. To
determine the correct Z factor, find the intersection between temperature and air pressure
in Table 6.3
6.3. Round up temperature and air pressure values.
Multiply each weight
m
i
obtained in 5.5.6 with the corresponding Z factor:
Add together the volumes
V
i
delivered, divide the sum by n (e.g. n = 5) to calculate the
mean volume
V (in millilitres or micro litres) delivered at the test temperature, which is the
Actual Volume:
The following symbols are used throughout this text:
V
s
= Selected test or target Volume
m
i
= Measured Liquid Weight (g)
Z = Z factor, see 6.36.3
V
i
= Converted Volume (ml or µl)
V = Actual mean Volume (ml or µl)
n = Number of measurements
Tar ge t V
s
Accuracy
Precision
V
Actual
V
i
m
i
Z=
V
1
n
---
V
i
i 1=
n
=
EVOLVE Manual Pipettes – Operating instructions V00 Maintenance
28 INTEGRA Biosciences AG
Calculation of systematic error (accuracy)
The systematic error
e
s
can be calculated using the following equation with V
s
being the
selected test volume:
or in percent:
Calculation of random error (precision %)
To calculate the random error as the repeatability standard deviation s
r
, use the following
equation:
The random error can also be expressed as a percentage, by the coefficient of variation
CV, using equation:
If the pipette is properly calibrated, the Target volume
V
s
should equal the Actual volume
V within the accuracy specifications of the pipette.
5.5.8 Adjusting EVOLVE Manual Pipettes
Compare the calculated accuracy and precision values with the corresponding pipette
specifications given in section 6.2
.
The industry standard is to test and present specifications using neat transfers. This is
aspirating and dispensing the same volume.
In case a EVOLVE Manual Pipette has not met the calibration specifications it needs to be
adjusted. This can be corrected with the Multipurpose Tool.
e
s
VV
s
=
e
s
100 VV
s
V
s
-----------------------------------
=
s
r
ViV–
2
i 1=
n
n 1–
-------------------------------=
CV 100
s
r
V
----
=
EVOLVE Manual Pipettes – Operating instructions V00 Maintenance
www.integra-biosciences.com 29
Put the two teeth of the Multipurpose Tool into the slots on the top of the pipette.
N
OTE
Validate the new volume set by repeating the gravimetric testing.
c)
5.6 Equipment disposal
The EVOLVE Manual Pipettes must not be disposed of with unsorted municipal
waste.
Dispose of the EVOLVE Manual Pipettes in accordance with the laws and
regulations in your area.
Rotate clockwise to increase the volume (slight turn up to a full turn).
Rotate counterclockwise to decrease the volume (slight turn up to a
full turn).
EVOLVE Manual Pipettes – Operating instructions V00 Technical Data
30 INTEGRA Biosciences AG
6 Technical Data
6.1 Environmental conditions
6.2 Pipette specifications
Precision = Coefficient of Variation.
Operation
Temperature range 5–40 °C
Humidity range Max. rel. humidity 80% for temperatures up to 31°C,
decreasing linearly to 50% rel. humidity at 40°C.
Altitude range <2000 m
EVOLVE Manual Pipettes Manufacturer
Chan-
nel
Part
No.
Volume Ran ge
(µl)
Volume
Increments
(µl)
Test
Volume
(µl)
Accuracy
(±%)
Precision
(≤%)
1 3011 0.2–2 0.001 0.2 12 6
12.81.6
21.8 1
1 3012 1–10 0.01 1 2.5 1.8
51.50.8
10 1 0.5
1 3013 2–20 0.01 2 5 2
10 1.6 0.5
20 1 0.35
1 3015 10– 100 0.1 10 3 1
50 0.8 0.35
100 0.8 0.2
1 3016 20– 200 0.1 20 2.2 0.9
100 0.8 0.3
200 0.7 0.18
1 3018 100– 1000 110030.6
500 0.8 0.3
1000 0.8 0.2
1 3019 500– 5000 5 500 2.2 0.7
2500 0.8 0.3
5000 0.6 0.18
EVOLVE Manual Pipettes – Operating instructions V00 Technical Data
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The multichannel pipettes’ specifications represent the performance of all channels
of the pipettes.
Precision = Coefficient of Variation.
EVOLVE Manual Pipettes Manufacturer
Chan-
nel
Part
No.
Volume Ran ge
(µl)
Volu me
Increments
(µl)
Test
Volume
(µl)
Accuracy
(±%)
Precision
(≤%)
83022 1–10 0.01 1 8.5 7
543
10 2 1.2
83024 5–50 0.05 5 4 2
25 2 1
50 1.2 0.5
83025 10–100 0.1 10 3.75 2
50 1.7 0.8
100 1.2 0.5
83026 20–200 0.1 20 4 1.3
100 1.5 0.5
200 1.2 0.4
12 3032 1–10 0.01 1 8.5 7
543
10 2 1.2
12 3034 5–50 0.05 5 4 2
25 2 1
50 1.2 0.5
12 3035 10– 100 0.1 10 3.75 2
50 1.7 0.8
100 1.2 0.5
12 3036 20– 200 0.1 20 4 1.3
100 1.5 0.5
200 1.2 0.4
EVOLVE Manual Pipettes – Operating instructions V00 Technical Data
32 INTEGRA Biosciences AG
6.3 Z Correction Factors
Z values in microliters per milligram
Temp.
(°C)
Air Pressure (kPa)
80 85 90 95 100 101.3 105
18.0
18.5
1.0022
1.0023
1.0023
1.0024
1.0023
1.0024
1.0024
1.0025
1.0025
1.0025
1.0025
1.0026
1.0025
1.0026
19.0
19.5
1.0024
1.0025
1.0025
1.0026
1.0025
1.0026
1.0026
1.0027
1.0026
1.0027
1.0027
1.0028
1.0027
1.0028
20.0
20.5
1.0026
1.0027
1.0027
1.0028
1.0027
1.0028
1.0028
1.0029
1.0028
1.0029
1.0029
1.0030
1.0029
1.0030
21.0
21.5
1.0028
1.0030
1.0029
1.0030
1.0029
1.0031
1.0030
1.0031
1.0031
1.0032
1.0031
1.0032
1.0031
1.0032
22.0
22.5
1.0031
1.0032
1.0031
1.0032
1.0032
1.0033
1.0032
1.0033
1.0033
1.0034
1.0033
1.0034
1.0033
1.0034
23.0
23.5
1.0033
1.0034
1.0033
1.0035
1.0034
1.0035
1.0034
1.0036
1.0035
1.0036
1.0035
1.0036
1.0036
1.0037
24.0
24.5
1.0035
1.0037
1.0036
1.0037
1.0036
1.0038
1.0037
1.0038
1.0037
1.0039
1.0038
1.0039
1.0038
1.0039
EVOLVE Manual Pipettes – Operating instructions V00 Accessories and Consumables
www.integra-biosciences.com 33
7 Accessories and Consumables
7.1 Accessories
7.2 Consumables
General Part No.
EVOLVE Pipette Multipurpose Tool
(for calibration, o-ring removal and plunger spring exchange)
3200
EVOLVE Pipette Shelf Hook, 1/pack 3210
EVOLVE Pipette Shelf Hook, 5/pack 3211
Linear Stand for manual/electronic pipettes 3215
Replacement Plunger Caps (5/pack) Part No.
Black for use with any pipette 3220
Purple for use with 2 µl, 10 µl, 20 µl manual pipettes 3221
Yellow for use with 50 µl, 100 µl manual pipettes 3222
Green for use with 200 µl manual pipettes 3223
Blue for use with 1000 µl manual pipettes 3224
Orange for use with 5000 µl manual pipettes 3225
EVOLVE Control Spring Assembly Part No.
for single channel pipettes 2, 10, 20, 100, and 200 µl only 3230
O-rings for tip fittings Part No.
200 μl,
(300 μl)
Replacement kit 24/pack 100-00027-50
1000 μl,
(1250 μl)
Replacement kit 24/pack 100-00028-50
5000 μl Replacement kit 10/pack 100-00029-00
Lubricant for pipettes and O-rings Part No.
Lube Parker Super-O-Lube, Silicone based, 50 g / 2 oz, for single
channel 1250 µl and 5000 µl pipettes
100-00135-50
Grease Grease Nye Fluorocarbon Gel 807, 5 g / 0.2 oz, for single
channel 12.5 µl, 125 µl and 300 µl pipettes
100-00136-50
EVOLVE Manual Pipettes – Operating instructions V00 Accessories and Consumables
34 INTEGRA Biosciences AG
GripTips for all INTEGRA Manual and Electronic Pipettes Part No.
Purple
LONG:
2 μl,
10 μl,
(12.5 μl),
20 μl
Bulk pack, 1 bag of 1000 tips, non-sterile, LONG 4401
5 inserts of 384 tips, non-sterile, LONG, GREEN CHOICE 4402
5 racks of 384 tips, non-sterile, LONG 4403
5 racks of 384 tips, sterile, LONG 4404
5 racks of 384 tips, sterile, filter, LONG 4405
5 inserts of 384 tips, pre-sterilized, LONG, GREEN CHOICE 4406
Purple:
2μl,
10 μl,
(12.5 μl),
20 μl
Bulk pack, 1 bag of 1000 tips, non-sterile 4411
5 inserts of 384 tips, non-sterile, GREEN CHOICE 4412
5 racks of 384 tips, non-sterile 4413
5 racks of 384 tips, sterile 4414
5 racks of 384 tips, sterile, filter 4415
5 inserts of 384 tips, pre-sterilized, GREEN CHOICE 4416
Yellow:
50 μl,
100 μl,
(125 μl)
Bulk pack, 1 bag of 1000 tips, non-sterile 4421
5 inserts of 384 tips, non-sterile, GREEN CHOICE 4422
5 racks of 384 tips, non-sterile 4423
5 racks of 384 tips, sterile 4424
5 racks of 384 tips, sterile, filter 4425
5 inserts of 384 tips, pre-sterilized, GREEN CHOICE 4426
Green:
200 μl,
(300 μl)
Bulk pack, 1 bag of 1000 tips, non-sterile 4431
5 inserts of 96 tips, non-sterile, GREEN CHOICE 4432
5 racks of 96 tips, non-sterile 4433
5 racks of 96 tips, sterile 4434
5 racks of 96 tips, sterile, filter 4435
5 inserts of 96 tips, pre-sterilized, GREEN CHOICE 4436
Blue:
1000 μl,
(1250 μl)
Bulk pack, 1 bag of 500 tips, non-sterile 4441
5 inserts of 96 tips, non-sterile, GREEN CHOICE 4442
5 racks of 96 Tips, non-sterile 4443
5 racks of 96 Tips, sterile 4444
5 racks of 96 Tips, sterile, filter 4445
5 inserts of 96 tips, pre-sterilized, GREEN CHOICE 4446
EVOLVE Manual Pipettes – Operating instructions V00 Accessories and Consumables
www.integra-biosciences.com 35
Orange:
5 ml
Bulk pack, 1 bag of 250 tips, non-sterile 4451
5 racks of 48 Tips, non-sterile 4453
5 racks of 48 Tips, sterile 4454
5 racks of 48 Tips, sterile, filter 4455
Individually wrapped, 100 tips per case, sterile 4456
GripTips for all INTEGRA Manual and Electronic Pipettes Part No.
EVOLVE Manual Pipettes – Operating instructions V00
36 INTEGRA Biosciences AG
Imprint
© 2016 INTEGRA Biosciences AG
All rights to this documentation are reserved. In particular the rights of reproduction,
processing, translation and the form of presentation lie with INTEGRA Biosciences AG.
Neither the complete documentation nor parts thereof may be reproduced in any way, or
stored and processed using electronic media or distributed in any other way without the
written consent of INTEGRA Biosciences AG.
This operating instruction manual has part number 900-00010-00, the version is V00.
EVOLVE and GripTip are trademarks of INTEGRA Holding, Switzerland.
Manufacturer
Customer service
Please contact your local INTEGRA Biosciences representative.
To find out name and address of your local representative go to
www.integra-biosciences.com
.
Further information and operating instructions in other languages are available on
www.integra-biosciences.com
or on request info@integra-biosciences.com.
INTEGRA Biosciences AG
CH-7205 Zizers, Switzerland
T +41 81 286 95 30
F +41 81 286 95 33
INTEGRA Biosciences Corp.
Hudson, NH 03051, USA
T +1 603 578 5800
F +1 603 577 5529
info@integra-biosciences.com
www.integra-biosciences.com
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