MICROLIT NERO User manual

TABLE OF CONTENTS:
1. Product Description
2. Product Features
3. Raw Material
4. Micropipette Operation
5. Tip Ejection
6. Pipetting Technique
7. Calibration
8. Micropipette Specifications
9. Error Calculation
14. Device Requirement and
Test Conditions
10. Maintenance
11. Short Term Service
12. Long Term Service
15. Performance Optimization
16. Autoclaving
17. Troubleshooting
18. Package
19. Caution
TABLE OF CONTENTS
1
1
2
2
2
3
5
6
7
7
7
7
11
12
12
13
14
8
13. Disassembly & Re-assembly
of Micropipette
10

Microlit NERO is made of mechanically durable
and autoclavable materials. It does not contain
any hazardous material.
1. Set the delivery volume
using the push button on the
top of the micropipette.
To decrease the delivery
volume, turn it clockwise.
Variable Volume:
Fixed Volume:
Raw Materials
Micropipette Operation
Easy tip ejection:
An in-built tip-ejector is designed
to eject tips with optimal force.
To increase the delivery
volume, turn the push button
counter-clockwise.
2. Make sure that the desired delivery volume
clicks into place.
3. Do not set a volume that lies
outside the micropipette’s
specified volume range.
There is no need to set the volume and the user
can directly operate the pipette as per the volume
required.
Product Description
MICROLIT NERO micropipettes are designed with
ergonomic and intuitive handling, ensuring high
precision with its µAir™ technology.
Built by our in-house team of product design
engineers, NERO Micropipettes are available in
Single Channel Fixed and Variable Volume ranges.
Product Features
Universal Tipcone:
Our tip-cone is designed to be compatible
with most internationally accepted tips,
which enhances the compatibility of the
instrument in the industry.
Microlit NERO offers sophisticated features
and it is designed to optimize the accuracy and
precision during operation.
µAir™ Technology:
The design of Microlit NERO
ensures minimum dead air
space between the piston and
the liquid in the tip, thereby,
reducing air compression and
ensuring high precision.
Tip Ejection
The pipette has a tip ejector
button which prevents
contamination. In order to
Minimum Dead Space
It is a unique calibration tool /
mechanism which allows quick
in-lab calibration in a single
operation Without disengaging
the digits from the plunger
mechanism.
Easy Volume Adjustment:
The easy rotation mechanism
of plunger drives the digital
system to set the volume of the
instrument comfortably.
Set the volume with perfection:
A soft click at every volume change
ensures perfect volume setting and
prevents any accidental volume
change.
A fixed volume micropipette
minimizes the random error as the
chances of accidental volume
change is nil. The fixed volume
allows the user to work at a faster pace.
eject the tip, point the micropipette at a suitable
waste receptacle and press the ejector button
with your thumb.
Note: Using excessive force to turn the push button
outside the range may jam the mechanism and
eventually damage the micropipette.
UniCal™ Technology (Patent Pending):
1 2

Forward Technique:
1. To aspirate the liquid in the
tip, press the plunger to the
first stop and Immerse the
pipette tip vertically in the
liquid.
2. Slowly release the plunger while the tip is
immersed. The liquid will be aspirated into the
pipette tip.
3. To dispense the liquid,
place the tip on the inner wall
of the receiving vessel at a steep
angle.
4. Slowly press the plunger to
the first stop to dispense the
liquid.
5. To empty the tip completely, press the plunger
to the second stop.
6. Wipe the tip on the inner wall while taking the
tip out of the vessel.
Reverse Technique:
1. To aspirate the liquid in the tip, press the
plunger to the second stop and immerse the
pipette tip vertically in the liquid.
2. Slowly release the plunger while the tip is
immersed. The liquid will be aspirated into the
pipette tip.
3. To dispense the liquid, place the tip on the
inner wall of the tube at a steep angle.
4. Slowly press the plunger to the first stop.
5. Wipe the tip on the inner wall while taking the
tip out of the vessel.
Note: Residual liquid remains in the tip.
This does not belongs to the dispense volume.
Repetitive Technique:
The repetitive technique offers a rapid and
sample procedure for repeated delivery of the
same volume.
1. To aspirate the liquid, press the plunger to the
second stop and immerse the pipette tip vertically
in the liquid.
2. Slowly release the plunger while the tip is
immersed. The liquid will be aspirated into the
pipette tip.
3. To dispense the liquid, place the tip on the
Inner wall of the vessel at a steep angle.
4. Slowly press the plunger to the first stop.
Hold the plunger at the first stop.
5. The residual liquid remains in the tip.
This does not belong to the dispense volume.
6. Continue pipetting by repeating the
steps 2 to 4.
Pipetting of Heterogeneous samples:
Let’s assume that we have to determine the
deproteinization in blood glucose.
1. Use the forward technique to fill the tip with
blood. Wipe the tip carefully with a dry and
clean tissue.
Pipetting Technique
General Instructions
1. Press and release the plunger slowly, at all
times, particularly when working with high viscosity
reagents / solutions.
Make sure that the plunger does not snap.
2. Make sure the tip is firmly attached to the tip
cone.
3. Before starting your experiment, fill and empty
the tip 2-3 times with the reagent or solution that
you will be pipetting.
4. Hold the micropipette
in an upright position while
aspirating.
The Grippy must rest on
your index finger.
5. Make sure that the tips, the micropipette
and the reagent/solution are at the same
temperature.
3 4

Calibration
Nut
Plunger
Calibration
Microlit NERO features UniCal™ Technology,
which allows the user to calibrate the pipette
without disassembling any component of the
pipette or digit disengagement or without
matching the calibration markers with the
adjustment chart in the operation manual.
The calibration tool is easy to use and enables
the user for quick in-lab calibration.
If the dispensed volume of the micropipette
(as read by an analytical balance) is not within
permissible ISO 8655 limits (as given in
Micropipette Specifications).
The pipette can be calibrated using the
calibration tool provided in the box by following
the procedure below:
1. Perform at least 5 measurements on the
nominal volume of the pipette and calculate
the average weighted volume as displayed
by an analytical balance.
2. Engage the calibration tool with the plunger
and the calibration nut and adjust the digits
to display the average weighted volume.
3. Once the average weighted volume
is displayed on the digital system,
remove the calibration tool and store
it safely. The pipette has now been
recalibrated.
Single Channel Variable Volume Micropipettes
The Single Channel Fixed Volume Micropipette
is available in 15 unique volume ranges from
1.0 ul to 10 ml.
Micropipette Specifications
The Single Channel Variable Volume
Micropipette is available in 9 unique volume
ranges from 0.2 ul to 10 ml.
Single Channel Fixed Volume Micropipettes
2. Immerse the tip into the reagent/solution and
press the push button till the first stop. Make
sure the tip is well below the surface.
3. Release the push button slowly to let it retract
to the ready position. This will fill the tip.
Keep the tip in the solution.
4. Press till the first stop and release slowly. Keep
repeating this procedure until the interior wall
of the tip is clear.
5. Finally, depress the push button all the way to
completely empty the tip.
6. Once the operation is complete, make sure
to eject the tip by pressing
the tip ejector.
Discard the used tip in
a bin to avoid any
contamination.
4. Adjust the volume back to the nominal
volume of the pipette by rotating the plunger.
5. Perform a few pipetting operations to check
the resulting volume. If the readings on the
balance are still outside permissible limits,
please perform the calibration procedure again.
Calibration Tool
5 6
NRO-2
NRO-10
NRO-20
NRO-50
NRO-100
NRO-200
NRO-1000
NRO-5000
NRO-10000
0.2-2.0 µl
0.5-10 µl
2-20 µl
5-50 µl
10-100 µl
20-200 µl
100-1000 µl
0.5-5 ml
1-10 ml
0.01
0.05
0.1
0.2
0.5
1.0
5.0
20.0
50.0
2
1
0.8
0.8
0.6
0.6
0.6
0.6
0.6
0.04
0.1
0.16
0.4
0.6
1.2
6
30
60
1.2
0.5
0.4
0.4
0.2
0.2
0.2
0.2
0.2
0.024
0.05
0.08
0.2
0.2
0.4
2
10
20
Model Volume
Range
Inc
µl
A CV
±% ±µl ±% ±µl
NRO-F-1
NRO-F-2
NRO-F-5
NRO-F-10
NRO-F-20
NRO-F-25
NRO-F-50
NRO-F-100
NRO-F-200
NRO-F-250
NRO-F-500
NRO-F-1000
NRO-F-2000
NRO-F-5000
NRO-F-10000
1.0
2.0
5.0
10.0
20.0
25.0
50.0
100.0
200.0
250.0
500.0
1000.0
2000.0
5000.0
10000.0
5
4
2
1
0.8
0.8
0.8
0.6
0.6
0.6
0.6
0.6
0.6
0.6
0.4
Model Volume ACV
±% ±µl ±% ±µl
0.05
0.08
0.1
0.1
0.16
0.2
0.4
0.6
1.2
1.5
3
6
12
30
40
(µl)
5
2
1
0.5
0.4
0.4
0.4
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.05
0.04
0.05
0.05
0.08
0.1
0.2
0.2
0.4
0.5
1
2
4
10
20

Disassembly & Re-assembly of Micropipette
A1. Disassembly:
(For single channel variable volume and
fixed volume 0.2 µl to 1000 µl)
Step 1: Rotate the tip ejector assembly
counter-clockwise to separate it from
the upper housing.
Step 2: Rotate the piston assembly counter-
clockwise to separate the piston from the
tip ejector housing.
Step 3: Pull out the o-ring seat and turn the
tip cone upside down and retrieve the o-ring.
Step 4: Clean the tip cone
Step 5: Grease the cleaned parts with a
lubricant, preferably silicon grease.
A2. Reassembly:
(For single channel variable volume and
fixed volume 5 ml and 10 ml)
Step 1: Place the o-ring in the tip cone and
screw the o-ring seat.
Maintenance
When the micropipette is not in use, make
sure that it is stored in an upright position.
We recommend a stand for this purpose.
Short Term Service
The micropipette must be checked at the
beginning of each day for dust and dirt on
the outer surface. Particular attention must
be paid to the Tip Cone.
No other solvents except 70% ethanol
must be used to clean the micropipette.
Long Term Service
If micropipette is used daily, it must be
checked every three months.
The servicing procedure starts
with its disassembly.
Error Calculation
Inaccuracy (Systematic Error):
Inaccuracy is the difference between
the dispensed volume and the
selected volume of a pipette.
_
A = V - V0
where,
A = Accuracy
_
V = Mean Volume
V0 = Nominal Volume
Inaccuracy can be expressed
as a relative value:
A% = 100% x A/V0
Imprecision (random error):
Imprecision refers to the repeatability of the
pipetting. It is expressed as the standard
deviation (S) of coefficient of variation (CV).
where,
S = Standard Deviation
_
V = Mean Volume
n = Number of measurement
Reproducibility or coefficient of variation (CV)
can be expressed as:
_
CV = 100% x S / V
7 8

Step 2: Place the spring and the o-ring seat
on the piston and place it inside the tip cone.
Step 3: Screw the
piston clockwise
in the tip cone.
Step 4: Screw the tip
ejector assembly in
the clockwise direction
to the upper housing.
B1. Disassembly:
(For single channel variable volume and fixed
volume 5 ml and 10 ml)
Step1: Rotate the tip ejector assembly counter-
clockwise to separate it from the upper housing.
Step 2: Pull the lower ejector housing to separate
it from the upper ejector housing.
Step 3: Rotate the tip cone in the counter-clockwise
direction to separate it from the piston.
Step 4:Grease the cleaned parts with a lubricant,
preferably Silicon grease.
B2. Reassembly:
(For single channel variable volume and fixed
volume 5 ml and 10 ml)
Step1: Screwback the tip cone to the piston
assembly by rotating it in the clockwise direction.
Step 2: Place the upper ejector housing and the
lower ejector housing on the piston assembly and
snap them back into place.
Step 3: Assemble the ejector housing to the upper
housing by rotating in the clockwise direction.
Device Requirements and Test Conditions
Use an analytical balance. The scale graduation
value of the balance must be chosen according
to the selected test volume of the micropipette.
The test liquid water is distilled or deionized
“grade 3” water, conforming to ISO 3696.
All the tests are done in a draft-free room
at a constant (±0.5°C) temperature of water
pipette and air between 15°C to 30°C.
The relative humidity must be above 50%.
For volumes under 50 μl, the air humidity
must be as high as possible to reduce the
effect of evaporation.
Special accessories, such as the evaporation
trap, are recommended.
Readable Volume Range
Pipette Graduation
under 10 μl
under 100 μl
Above 100 μl
Balance Graduation
0.001 mg
0.01 mg
0.1 mg
9 10

Performance Optimization
Activity
Consistent Technique Pipette with a consistent
rhythm, pressure
and speed.
Tip Size and Fit Use proper size tip, firmly
placed on Tip Cone.
Action
Pre-rinse Tips Pre-rinse pipette tips for
improved precision.
Sample Aspiration Keep the disposable tip
immersed in fluid during
aspiration. Do not let the
plunger snap back to
starting position.
Immersion Depth Maintain an immersion
depth of 2 to 4 mm.
Viscous Samples Aspirate slowly. If bubbles
are observed, resample.
Volume errors may still
occur. Refer to the
Calibration section.
Acid Samples
High Vapor
Pressure Samples
Sample Temperature Fluids at a temperature
other than that for which
the pipette and pipette
tips have been calibrated
may result in volume
measurement errors.
Storage Store upright in stand.
Do not lay the pipette
on its side with fluid in
the tip. Fluid reaching
the piston causes
contamination and
possible corrosion.
Cleaning Piston Wipe piston with alcohol
and a soft, lint-free cloth.
Dry and lightly lubricate
the piston.
Performance Checks Check the accuracy and
precision of your pipette
every 3-6 months
depending on use and
the samples aspirated.
Pipetting solutions with
high vapor pressure is
not recommended.
These liquids may damage
the piston and seal.
Pipetting strong acids
and corrosive solutions
is not recommended.
These liquids may damage
the piston and seal.
Autoclaving
• The micropipettes can be sterilized by
autoclaving them at 121°C (252°F) at 15 psi
for a minimum of 20 minutes.
• No special preparation is needed.
• You may use steam sterilization bags
if needed. After autoclaving, the micropipette
must be cooled to room temperature for at
least two hours. Before pipetting, make sure
that the instrument is dry.
• We recommend that you check the calibration
after every autoclave cycle to achieve the
best possible precision and accuracy.
Troubleshooting
The table below lists possible problems
and their solutions.
Problem
Liquid is leaking
from pipette tip.
Amount of
sample delivered
is not accurate.
Possible Reason
Pipette tip does
not fit properly
onto the tip cone.
Proposed Action
Use the
appropriate tip
for exact tip
fitment.
Liquid being
pipetted is hot
or cold. Liquid
being pipetted
is very dense
or viscous.
Shorten the
amount of time
the liquid is in
the tip, or the
possible solution
is to use the
reverse mode
of pipetting.
Pipette sealing
O ring is worn. Replace the sealing O ring.
Tip is not loaded
to maintain
the seal.
Press on tip
more firmly.
(Avoid using too
much force and
over inserting
the tip)
Pipette is not
within calibration
specifications.
Re-calibrate the
pipette as per
the operation
Manual.
Improper pipette
technique.
Foreign particles
between tip and
Tip Cone.
Clean the Tip
Cone with a lint
free cloth and
attach new tips.
Refer pipetting
technique.
11 12

Package
The micropipettes are shipped in
specially designed packages
containing the following items.
Caution
The micropipettes are designed to allow
easy in-lab service. If you would prefer to
have us or your local representative service
your instrument, please make sure it has
been decontaminated before you send
it to us.
Please note that the postal authorities
in your country may prohibit or restrict the
shipment of contaminated material by mail.
Calibration Tool
Tip Sample
The Micropipette
Shelf Hanger
Operation Manual
Calibration Certificate
13 14