Malvern NANOSIGHT NS300 User manual

NANOSIGHT NS300
OPERATING MANUAL
MAN0516-05-EN-00 September 2014

NanoSight NS300 Operating Manual
(MAN0516-05-EN-00) 2
Copyright © 2014 Malvern Instruments Ltd.
Malvern Instruments pursues a policy of continual improvement due to technical development. We therefore reserve the
right to deviate from information, descriptions, and specifications in this publication without notice. Malvern Instruments
shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing,
performance or use of this material. No reproduction or transmission of any part of this publication is allowed without the
express written permission of Malvern Instruments Ltd.
Head office:
Malvern Instruments Ltd.
Enigma Business Park,
Grovewood Road,
Malvern,
Worcestershire WR14 1XZ
United Kingdom.
Tel + [44] (0)1684-892456
Fax + [44] (0)1684-892789
www.malvern.com
NanoSight, Malvern and the 'hills' logo are registered trademarks in the UK and/or other countries, and are owned by
Malvern Instruments Ltd.

NanoSight NS300 Operating Manual
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NANOSIGHT NS300
OPERATING MANUAL
Contents
1.1 INTRODUCTION ......................................................................................................................................... 5
1.2 EXPLANATION OF WARNING SYMBOLS ................................................................................................. 5
1.3 IMPORTANT NOTES .................................................................................................................................. 5
1.3.1 SAFETY ................................................................................................................................................... 5
1.3.1.1 Laser Safety ................................................................................................................................... 5
1.3.1.2 Electrical Safety.............................................................................................................................. 6
1.3.1.3 General Safety ............................................................................................................................... 6
1.3.2 SERVICING ............................................................................................................................................... 7
1.3.3 MAINTENANCE ......................................................................................................................................... 7
1.3.4 HANDLING............................................................................................................................................... 7
1.3.5 UNPACKING AND INITIAL INSPECTION............................................................................................................. 7
1.3.6 INSTALLATION AND RELOCATION .................................................................................................................. 8
1.3.7 RETURNING EQUIPMENT.............................................................................................................................. 8 
1.3.8 WARRANTY ............................................................................................................................................. 8
2. NANOSIGHT NS300 HARDWARE................................................................................................................ 9
2.1 NS300 TECHNICAL SPECIFICATIONS ........................................................................................................ 9
2.1.1 NS300 INSTRUMENT HOUSING.................................................................................................................... 9
2.1.2 NS300 LASER MODULE ............................................................................................................................. 9
2.1.3 LASER CLASSIFICATION ............................................................................................................................... 9
2.2 NS300 OPERATION.................................................................................................................................. 10
2.2.1 INSTRUMENT SET-UP ................................................................................................................................ 10
2.2.2 DAILY USE............................................................................................................................................. 10
2.2.3 LASER MODULE CONFIGURATION ............................................................................................................... 11
2.2.3.1 Flow-cell top-plate ....................................................................................................................... 12
2.2.3.2 O-ring top-plate........................................................................................................................... 13
2.2.4 ASSEMBLING AND CLEANING THE LASER MODULE.......................................................................................... 15
2.2.4.1 Flow-cell top-plate ....................................................................................................................... 15
2.2.4.2 O-ring top-plate........................................................................................................................... 17
2.2.5 LOADING A SAMPLE................................................................................................................................. 18
2.2.5.1 Using the flow-cell top-plate........................................................................................................ 19
2.2.5.2 Using the O-ring top-plate ........................................................................................................... 19
2.2.5.3 Using the syringe-pump............................................................................................................... 19
2.2.6 MOUNTING THE LASER MODULE ................................................................................................................ 20
2.2.7 LOADING A NEW SAMPLE ......................................................................................................................... 20
2.2.7.1 Flow-cell top plate ....................................................................................................................... 20
2.2.7.2 O-Ring top plate .......................................................................................................................... 20

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3. NANOSIGHT NS300 CONTROL SOFTWARE ............................................................................................. 21
3.1 NTA SOFTWARE STARTUP............................................................................................................................ 21
3.2 CAMERA SETUP ......................................................................................................................................... 22
3.3 ADJUSTING THE NS300 TO OBTAIN A VIDEO .................................................................................................... 22
3.4 THRESHOLDING PIXELS (HIGH-SENSITIVITY SYSTEMS ONLY) .................................................................................. 23
3.5 FLUORESCENCE MODE ................................................................................................................................ 23
3.5.1 Viewing fluorescently labelled particles ........................................................................................... 23
3.6 CAPTURE VIDEOS ....................................................................................................................................... 24
3.7 TEMPERATURE CONTROL ............................................................................................................................. 24
3.7.1 Operation ....................................................................................................................................... 24
3.7.2 Initialising ....................................................................................................................................... 25
3.7.3 Temperature Setting ....................................................................................................................... 25
5. SPARE PARTS LIST ..................................................................................................................................... 27

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1.1 Introduction
The NanoSight NS300 instrument is a patented, laser-based, light scattering system which provides an easy-to-use,
reproducible platform for specific and general nanoparticle characterisation.
Particles suspended in a liquid are loaded into the laser module sample chamber and viewed in close proximity to the optical
element. The NS300 device illuminates the particles using a specially aligned and focussed laser beam. This allows extremely
small particles (down to 10 nm) to be seen directly and individually by conventional microscopy.
Particles in the liquid sample which pass through the beam path are seen as small points of light moving rapidly under
Brownian motion, allowing information on particle properties to be obtained. With the NS300 you can analyse the
presence, size distribution, concentration and fluorescence of all types of nanoparticles from 10 nm to 1000 nm, depending
on the instrument configuration and sample type.
The laser module contains thermoelectric Peltier elements, allowing the sample temperature to be controlled. This is fully
programmable using the NTA Software Suite.
The installed tubing and the viewing chamber surfaces allow the use of all non-flammable, water-based solvents with
neutral pH. Other solvents may not be compatible with wetted surfaces and should not be used without confirming
suitability. If in doubt about the choice of solvent and its compatibility with any part of the device, please contact Malvern
([email protected]) for further information.
This manual is designed to provide an introduction to help familiarise the user with the NanoSight NS300 operation. Further
advice and help to get the best out of your instrument can be found in the NanoSight Application Notes, Technical Notes,
NTA Software Operation Manual, Quick-start Guides and User Training Videos. These can be found on our website
(www.Malvern.com) or by contacting Malvern.
For technical assistance, please contact Malvern
Malvern Technical Support:Tel: + [44] (0)1684-892456 or Email: [email protected]
1.2 Explanation of Warning Symbols
Warning: read instructions to understand possible hazard prior to use.
Electric shock hazard
1.3 Important Notes
1.3.1 Safety
1.3.1.1 Laser Safety
•The NanoSight NS300 device is classified (to BS EN 60825-1 (2001)) as a Class 1 laser device. The instrument
contains a Class 3B laser which must never be removed from the laser module housing.
•The laser module should not be plugged into the main body of the NS300 without the top plate being secured in
position with the screws supplied.
•The laser module should always be removed from the main body of the NS300 (cutting power to the laser) prior to
removing the top-plate screws.

NanoSight NS300 Operating Manual
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6
1.3.1.2 Electrical Safety
• Fuses must be only replaced with a fuse of the type and rating - F5AH250V Ø5mm x 20mm long.
• Use only the power adapter and other accessories supplied with the instrument.
• If the mains cordset needs to be replaced, please select another cordset which conforms to the following
specifications:
Voltage Rating
125 V AC if being used with a 100-120 V supply.
250 V AC if being used with a 220-240 V supply.
Current Rating
6 A minimum.
Temperature Rating
60 °C minimum.
Length
3 m Maximum.
Fittings
Grounded plug for attachment to power outlet.
IEC appliance coupling.
In addition to the above specifications, the mains cordset should be certified by one of the following institutions.
UK – BSI USA – UL Europe – VDE Japan – JET, JQA, TUV
• If the appliance is being used outside of the above areas the local regulations for mains power cords must be
checked and a cordset which complies with the relevant standards should be sourced for use with the equipment.
• The safety of the equipment can no longer be warranted if a non-approved cordset is used.
1.3.1.3 General Safety
• The instrument must not be used in hazardous areas.
• The instrument is for use in moderate climates only. Never use the equipment in damp or wet conditions.
• Avoid excessive heat, humidity, dust and vibration.
• Do not place liquid filled containers on the equipment.
• Do not use where the equipment may be subjected to dripping or splashing liquids.
• If the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment
may be impaired.
• The instrument can be used with all non-flammable water-based solvents of neutral pH. Other solvents may not be
compatible and must not be used without first contacting Malvern to confirm suitability.
• When using bio or chemically hazardous sample material, it is the responsibility of the operator to determine the
requisite protection for each application.
• The NS300 system contains no user serviceable part. It should not be modified in any way. Any modification will
void the warranty and could make the device unsafe.
• Use of controls, adjustments or performance of procedures other than those specified herein may result in
hazardous radiation exposure.

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•The laser module may become hot when operating the temperature control. Caution should be taken when
handling the device.
•The temperature control should always be turned off when the instrument is unattended.
1.3.2 Servicing
•The NS300 must only be serviced by qualified Malvern personnel, or Malvern approved agents.
•The NS300 contains no user-serviceable parts. The instrument casing should not be opened by any user.
•Removing the instrument casing or opening the housing of the laser module voids all warranties and could expose
users to hazardous voltages or Class 3B laser radiation.

1.3.3 Maintenance
•The NS300 system contains no user serviceable parts. It should not be modified in any way. Any modification will
void the warranty and could make the device unsafe.
•ThecasingoftheNS300bodyandthelasermoduleshouldbekeptcleanwiththeuseofadampcloth.Donotwetor
allowexcessmoisturetopenetrateanypartofthesystem.Donotusesolvents.
•To maintain best functionality and to protect against cross contamination during and following use, the sample
flow cell and tubing should be cleaned as described in this manual.
•Regularly inspect fluidic tubing and replace any that show signs of wear. Please contact Malvern for additional sets
of tubing (P/N NTA4038 NS300 Tubing Kit).
1.3.4 Handling
The NanoSight NS300 incorporates a rugged housing designed to protect the integrity of the instrument. However, it is a
sensitive scientific instrument and should be treated as such. The laser module contains a static-sensitive laser diode and
should never be used in circumstances when a static discharge may damage the diode.
During use and when cleaning it is important to ensure that liquid does not enter the inside of the laser module. Use only a
damp tissue for cleaning, never expose the laser module to excess fluids. During sample loading and instrument operation,
ensure that the sample chamber is properly sealed and that there is no evidence of any leaking fluids.
Between uses of the NanoSight NS300, or for longer term storage, the unit must be cleaned and dried as described in this
manual.
1.3.5 Unpacking and Initial Inspection
The standard instrument is supplied with the following accessories:
•Mains power lead
•USB cable
•CMOS Firewire cable
•Spare NS300 Tubing Kit
•Spare gasket component
•Instruction manuals
•2mm Allen/Hex-key with flow-cell nut
Inspect the shipping container when the NS300 is received. Carefully check the contents for completeness and condition.
The NS300 is supplied in a specially designed packing case which should be returned to Malvern after delivery. The packing
case can be kept if required, although an additional charge will be made.
Notify Malvern ([email protected]) if the contents are incomplete, or if the instrument or accessories appear to be
damaged in any way. Keep all damaged packaging, materials and goods for inspection by the carrier.

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1.3.6 Installation and Relocation
Initial installation will be carried out by qualified Malvern personnel.
In the event that the instrument needs to be relocated the following instructions must be adhered to:
Always disconnect the equipment from the mains and ancillary units before moving.
The instrument should be located in an area of good ventilation and with sufficient space for safe and efficient
operation and maintenance.
The supplied PC should be sited to avoid dangers from spillage and splashing.
The equipment must be connected to an earthed power supply with a voltage corresponding to that on the power
adapter.
Ensure that the mains plug is easily accessible to allow the unit to be disconnected from the mains supply.
Always use the mains lead supplied. Your sales representative can provide a lead suitable for your country.
1.3.7 Returning Equipment
If, for any reason, you experience problems with your instrument, contact Malvern ([email protected]).
In the unlikely event you experience a problem with the NS300 system that requires returning the unit for repair, please
contact Malvern for instructions and documentation:.
The following information will need to be supplied:
•Sender’s name and address;
•Sender’s contact telephone number and email address;
•Complete list of equipment being returned including serial numbers;
•A detailed description of the problem or reason why the equipment is being returned;
•Declaration that, if the instrument has been used with bio or chemically hazardous sample material, all
equipment has been fully decontaminated before return.
On receipt of this information Malvern will provide a Return Merchandise Authorization (RMA) number. An RMA number
must be obtained from Malvern before returning any equipment, and should be clearly displayed on the return shipment
and included on all subsequent correspondence.
If the original shipping case is not available, shipping cases will be supplied by Malvern as required to ensure safe return of
the system
1.3.8 Warranty
Malvern warrants that the NS300 system as supplied with its accessories is free from defects in materials and workmanship
for a period of one year from shipping to the customer. During this warranty period, Malvern will, at its discretion, repair or
replace defective products.
Any liability under this warranty extends only to the replacement value of the equipment.
This warranty is void if:
•The NS300 or its accessories have been partly or completely disassembled, modified or repaired by
persons not authorised by Malvern.
•The instrument or instrument system is installed or operated other than in accordance with these
operating instructions.
No other warranty is expressed or implied. Malvern is not liable for consequential damages except as limited by English law.

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2. NanoSight NS300 Hardware
2.1 NS300 Technical Specifications
2.1.1 NS300 Instrument Housing
Size: 400 mm x 400 mm x 250 mm
Weight: ~12 kg
Operating temperature: 10 °C to 40 °C
Humidity: Up to 80% rH at 31 °C then decreasing linearly to 50% at 40 °C
Casing material: Plastic, aluminium
Voltage: 100-240 V rms +/- 10%
Frequency range: 50 – 60 Hz
Max power: 60 VA
2.1.2 NS300 Laser Module
Size: 140 mm x 74 mm x 68 mm
Weight: 650 g
Temperature control range: from 8 °C below ambient to 50 °C
Temperature control accuracy: +/-1 °C
Time to temperature: 3 minutes (to indication of within 1 °C)
Humidity: 5-95% non-condensing
Casing material: Anodised aluminium alloy
Top plate materials: Flow-cell top-plate: PMMA, glass, PDMS
SU8 epoxy, Norland Optical Adhesive 61
O-ring top-plate: Anodised aluminium alloy, glass and viton rubber
Wetted Parts: PTFE tubing
Flow-cell top-plate: PMMA, glass, PDMS, PEEK connectors
SU8 epoxy, Norland Optical Adhesive 61
O-ring top-plate: Anodised aluminium alloy, glass,
Viton rubber and Delrin connectors
2.1.3 Laser Classification
Embedded laser: Red 642 nm CW, max power < 50 mW
Green 532 nm CW, max power < 60 mW
Blue 488 nm CW, max power < 55 mW
Violet 405 nm CW, max power < 70 mW

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2.2 NS300 Operation
2.2.1 Instrument Set-up
1) The NS300 instrument communicates with the controlling PC via a USB
connection. Plug the USB cable into the USB port on the right side of the NS300
instrument casing (labelled USB-1) and connect it to one of the USB ports on the
supplied PC.
2) Plug the firewire camera cable into the firewire port on the side of the NS300
(labelled Firewire B) and connect it to the PC firewire port.
3) On the right-hand side of the NS300 casing is a tubing port holder. The tubing
can be used to introduce sample when using the flow-cell top-plate or a syringe
pump accessory. The holder can be slid in and out of the case for easy access to
the fluidic connectors. The tubing port holder is held in place with a black nylon
screw on the inside of the casing, accessed by opening the front hatch.
4) The left-hand port on the tubing holder is the inlet port, used to introduce
sample into the cell. The right-hand tubing port is the waste port:
i) For injection from a disposable syringe, connect the tubing line with luer
attachment to the left-hand port.
ii) For use with the syringe pump accessory, connect the sample inlet tubing
from the syringe pump to the left-hand port.
iii) Connect a length of waste tubing to the right-hand port and put the
other end in a suitable waste receptacle.
5) Set up and turn on the PC.
6) Connect the mains lead to the NS300 instrument and turn the instrument on
at the power switch, located on the left side of the case.
2.2.2 Daily Use
The NS300 should always be left clean and dry whenever the system is not in use. The NTA software should be shut down
and the NS300 power switch turned off.
After use, the system should be flushed through with clean diluent to remove all traces of sample from the tubing and
optical surfaces. If using diluent with a high concentration of dissolved solids (i.e saline solution), flush clean water through
the system after use.
After the fluidic system is clean, empty the top-plate by extracting all liquid with a syringe, and then manually disassemble,
clean and dry the sample chamber as described in Section 2.2.4.
Between uses of the instrument, the sprung fastening screws holding the top-plate down should be
fully loosened, so that the optical flat is not held under any pressure. Failure to loosen the top-plate
screws between uses can result in damage to the optical flat.

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2.2.3 Laser Module Configuration
The laser module contains a specially configured and focussed laser source, mounted within a sealed housing. An optical
flat element is fixed in place on the top cover of the housing. The electrical contacts on the end of the housing connect to
the contacts within the NS300 to provide the laser module with power.
Optical flat
Power contact
The laser module can be supplied with a choice of two different top-plates, as shown below:
The flow-cell top-plate, can be cleaned by flushing wash fluid through the chamber, avoiding the need to manually
disassemble and clean the chamber after every sample.
The O-ring top-plate requires manual cleaning after each sample, but provides greater chemical compatibility for
non-aqueous solvents, or samples which are more viscous or contain larger particles, which may block the flow-cell.
The sample chamber configuration and the instructions for cleaning will vary according to which design of top-plate is
being used. Please refer to Section 2.2.3.1 and 2.2.4.1 for the flow-cell top-plate and Section 2.2.3.2 and 2.2.4.2 for the O-
ring top-plate, as appropriate.
Flow-cell top plate O-ring top-plate

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2.2.3.1 Flow-cell top-plate
For the flow-cell top-plate, the NS300 fluidic tubing is connected to ports on the top-plate, which is mounted onto the
optical flat on the laser module to form a sample chamber.
The sample chamber is formed by two parts which are assembled and then attached on top of the optical flat on the laser
module.
A gasket component, with an embedded microchannel, is connected onto the flow cell top-plate support structure
using a fixing bolt, tightened using an Allen / Hex key.
Two circular seals on the underside of the top-plate support structure seal the gasket component onto the support
structure
The combined top-plate is then sealed onto the laser module using four sprung fastening screws
The gasket component seal is compressed onto the optical flat to make an enclosed sample chamber.
The top-plate support structure circular seals (Part No NTA4012 and the gasket component (Part No. NTA4057) can be
replaced if the seals become worn or damaged.
For instructions on how to clean and assemble the flow-cell top plate see Section 2.2.4.1
Flow cell Top-Plate
Sprung fastening
screws
(spring held by circlip
attached to the bolt)
Flow cell top plate
support structure
Fixing bolt
Gasket component
Sprung fastening
screws
Enclosed sample
chamber
NS300 fluidic
tubing
The tubing on the left-hand
port on the top-plate should
be attached to the left inlet
port on the casing of the
NS300.
The tubing on the right-hand
port of the top-plate should
be attached to the right waste
port on the casing of the
NS300.

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2.2.3.2 O-ring top-plate
The O-ring top-plate contains an embedded O-ring seal which is mounted onto the laser optical flat to form the sample
chamber. The top-plate is sealed onto the laser module using four sprung fastening screws. The O-ring top-plate can be
setup in two different ways, according to how samples are to be loaded into the system.
2.2.3.2.1 Direct syringe connection
Samples can be loaded into the chamber using a disposable syringe, directly connected to a luer port on the top-plate. A
push-fit elbow luer connector is provided so that the laser module fits into the NS300 with the syringe remaining in place.
Insert the disposable syringe into the front luer port for sample loading (see
Section 2.2.5).
Once the sample has been loaded, the laser module should be placed into
the NS300, keeping the syringe connected
For instructions on how to assemble and clean the O-ring top plate see
Section 2.2.4.2
O-ring Top-Plate
Sprung fastening
screws
(spring held by circlip
attached to the bolt)
O-ring seal
Sprung fastening
screws
Push-fit luer
connectors
Enclosed sample
chamber

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2.2.3.2.2 Fluidic tubing connection
A sample can also be loaded into the O-ring top-plate through the NS300 fluidic tubing when using a syringe pump
accessory. In this case, the luer ports should be removed and the tubing from the tubing port holder attached to the ports
on the top-plate, using the connectors provided. The syringe pump can then be connected externally to the tubing port
holder on the outside of the NS300 casing.
Sprung fastening
screws
(spring held by circlip
attached to the bolt)
O-ring seal
NS300 fluidic tubing
Enclosed sample
chamber
Sprung fastening screws
The outlet tubing (wider bore) should be attached to the back port of the top-
plate and to the right waste port on the casing of the NS300.
The inlet tubing (narrower bore) should be attached to the front port of the top-
plate and to the left inlet port on the casing of the NS300.
For instructions on how to assemble and clean the O-ring top plate see Section
2.2.4.2
Important!
The fluidic tubing connection is intended for use with the syringe pump accessory only. Manually loading sample through a
syringe with the waste tubing connected can exert high pressures on the chamber seal. This can result in sample leakage or
damage to the top-plate.

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2.2.4 Assembling and Cleaning the Laser Module
Whilst the unit is splash-proof, under no circumstances must excess liquid be allowed to enter the
housing of the laser module at any time. This will cause irreparable damage to the laser mounted
within the unit.
Between uses of the instrument, the sprung fastening screws holding the top-plate down should be
fully loosened, so that the optical flat is not held under any pressure. Failure to loosen the top-plate
screws between uses can result in damage to the optical flat.
It is important to take care not to abrade or scratch the optical flat surface, or introduce particulates or contaminants onto
the surface. Treat all optical surfaces with the same care as would be employed with equivalent surfaces on your
microscope. Should the optical flat in the laser module become damaged, it can be replaced by Malvern (pricing on
request).
The following paragraphs provide recommended guidelines for cleaning the laser module. If necessary, but dependent on
the top-plate in use, other cleaning agents may be employed such as dilute mild detergents, or even dilute acid washes
prior to a final solvent cleaning step. If in doubt about the choice of solvent and its compatibility with the top-plate, optical
flat or any other part of the NanoSight device, please contact [email protected]om for further information.
2.2.4.1 Flow-cell top-plate
The flow-cell top plate has been designed to not need manual cleaning between every sample. Between samples, push
water or diluent through the sample chamber, to remove any particles present.
A manual clean is only necessary if there is visible cloudiness or sample residue stuck to the optical flat or to the glass
window in the flow-cell top-plate.
The glass window in the top-plate is fragile, and should be treated with care during manual cleaning.
1) Before manual cleaning, flush water or cleaning solution through the sample
chamber.
2) Extract the liquid in the sample chamber using the inlet syringe, making sure that no
liquid is sucked back up from the waste bottle.
3) The cell can then be disassembled, cleaned and reassembled as follows.
i) Open the door on the NS300 instrument and rotate the red lever to the left to
release the laser module. Slide out the laser module from inside the NS300 so
that the sample chamber can be easily accessed. There is no need to remove
the tubing for the purposes of cleaning.
ii) Unscrew the metal screws and springs securing the top plate and lift the top
plate gently off the laser module. Important: Avoid touching the channel seal
on the bottom of the gasket component.
iii) Wet a tissue with water, or water then ethanol if needed, and use this to
wipe the optical flat on the laser module. Do not pour any liquid over the laser
module, as this could penetrate the casing and damage the laser inside. Wipe
the flat gently with a soft dry tissue (e.g. Mediwipes) to remove any streaks
from the optical surface and then dry with compressed air.

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iv) Avoiding touching the chamber seal on the bottom of the gasket component, undo the fixing bolt with the 2
mm Allen/Hex-key until it is loose enough to slide out the gasket component. Avoid completely unscrewing the
bolt
v) Using a dripping or low-pressure source, e.g. a water bottle, to avoid causing any damage, rinse the circular seals
on the underside of the top-plate support with water, or water then an up to 10% ethanol-water solution if
needed, to remove any sample particles.
vi) Rinse the seal and the glass on the underside of the
gasket component with a low-pressure source of
water, or water then an up to 10% ethanol-
water solution. Take care to limit any rubbing of
the channel seal surrounding the glass window,
If any sample residue is visible on the gasket
viewing window, the underside of the glass can be
wiped from the bottom of the gasket component
with a small cotton bud
Do not apply any pressure to the top surface of the glass, as this may cause the viewing window
to be pushed out and separated from the gasket component
vii) Direct compressed air through the embedded micro-channel via the holes on the top of the gasket component
(i.e. from the side without the attached seal) to dry the channel.
viii) Keeping the nozzle of the compressed air can at least 5 cm from the seal, lightly dry the
underside of the component and the glass surface section using compressed air.
Important: Avoid touching or rubbing the seal when drying the underside. Keep the
compressed air nozzle at least 5cm away from the seal, to avoid high pressure damaging it
Important: If the gasket component repeatedly leaks after a manual clean, it should be
replaced (Part No. NTA4027).
a) Dry channel thoroughly through the holes on the
top of the gasket component
b) Dry underside of the gasket component, keeping the
air duster nozzle away from the seal

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ix) When all parts of the top plate are clean and dry, reattach Fixing bolt nut
the gasket component to the top-plate support using the
fixing bolt. Use an Allen/Hex-key to tighten the bolt, just until
the bolt nut on the underside is fully recessed into the
gasket component with no protrusion. You should not be able
to feel the nut if you run your finger over it.
x) Place the top-plate back onto the optical flat on the laser
module. Position the top plate using the locating pins on
the laser module and the corresponding holes in the top plate.
xi) Place the four metal screws and springs into the screw holes. For each screw in
turn, gently push down each screw head, compressing the spring, and twist
clockwise until the just screw locks in place in the screw thread underneath.
Tighten the four screws evenly in diagonal pairs until all are finger-tight. Do not
use Allen/Hex-keys or over-tighten the screws as this may cause damage to the
screw threads or damage the optical flat.
xii) Ensure there are no kinks in the waste outlet capillary tubing - this can lead to an
increase in pressure in the chamber.
2.2.4.2 O-ring top-plate
For the O-ring top-plate, a manual clean is required between samples to ensure there is no carry-over.
i. If the top-plate is connected to the NS300 fluidic tubing and a syringe pump,
load a syringe of water or cleaning solution through the sample chamber and to
waste, to remove any sample from the tubing lines and the top-plate
connectors.
ii. Extract the liquid in the sample chamber using the inlet syringe, ensuring no
liquid is sucked back up from the waste bottle if the syringe pump tubing is
connected
iii. Open the door on the NS300 instrument and rotate the red lever to the left to
release the laser module. Slide out the laser module from inside the NS300 so
the sample chamber can be easily accessed. There is no need to remove any
tubing for the purposes of cleaning.
iv. Unscrew the sprung fastening screws securing the top plate and lift the top
plate gently off the laser module.
v. Rinse the top-plate with
water, or water then
ethanol if needed, including
the O-ring seal and inside the
fluidic ports.

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vi. Clean the inside of the window by gently wiping it with a tissue wet with water,
or water then ethanol if needed, to remove all traces of the sample being
analysed.
vii. Wipe the glass window of the top plate gently with a soft dry tissue (e.g.
Mediwipes) to remove any surface streaks, then dry with compressed air.
viii. Wet a tissue with water,or water then ethanol if needed, and use this to wipe
the optical flat on the laser module. Do not pour any liquid over the laser
module, as this could penetrate the casing and damage the laser inside. Wipe the
flat gently with a soft dry tissue to
remove any streaks from the optical surface, then dry with compressed air.
ix. When all optical surfaces are clean and dry, place the top-plate back onto the
laser module. Position the top plate using the locating pins on the laser module
and the corresponding holes in the top plate.
x. Place the four sprung fastening screws into the screw holes. For each screw in
turn, gently push down each screw head, compressing the spring, and twist
clockwise until the screw locks in place in the screw thread underneath. Tighten
the four screws evenly in diagonal pairs until all are finger-tight. Do not over-
tighten the screws as this may cause damage to the screw threads or damage the
optical flat.
xi. If using the waste outlet tubing ensure there are no kinks - this can lead to an
increase in pressure in the chamber.
2.2.5 Loading a Sample
Once the top-plate is in place and any tubing connected, the system is
ready to load a sample. We recommend that before proceeding with
analysis, a sample of any buffer or diluent is checked to confirm that it
doesn’t contain any contaminating nanoparticles.
For both top-plates, the first sample should be loaded with the laser
module outside of the instrument. As the sample is loaded, the presence
of any air pockets or bubbles can be detected and removed.
Sample can be loaded into the chamber with a syringe connected to the
inlet tubing for the flow-cell top-plate, or directly connected to the luer
fitting on the O-ring top-plate. Samples can also be loaded using a
syringe pump accessory connected to the inlet tubing.
Important!
Occasionally a bubble may be present in the chamber and in the path of the laser beam. This might cause some
degree of specular reflection of the laser beam off the bubble surface. This will degrade image quality and should
be removed before analysis. Certain samples may generate bubbles through out-gassing of dissolved gases over extended
periods. In both cases, the image collected by the camera supplied will show a high intensity region (or may be completely
saturated) indicative of the presence of such bubbles. These can sometimes be removed by carefully moving fresh sample
through the chamber, allowing air bubbles to disperse. If samples repeatedly show evidence of bubble formation, it is
advisable to de-gas the sample before analysis.

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2.2.5.1 Using the flow-cell top-plate
i. Fill a syringe (1 ml to 2.5 ml) with the appropriate sample
Note: large bore syringe bodies (>5 ml) can inadvertently produce high pressures which may damage the top-plate
and should not be used.
ii. Remove any air bubbles from the syringe
iii. Place the syringe into the luer port on the end of the inlet tubing
iv. Introduce the sample slowly into the chamber.
Note: When loading the sample cell from dry, liquid should be introduced as slowly as possible, i.e. 1ml in ~10
seconds. This avoids generating pressures which might result in the sample bypassing the seals in the sample
chamber or damaging the top-plate. Loading the sample slowly also limits the introduction of bubbles.
2.2.5.2 Using the O-ring top-plate
Important!
The laser module should always be removed from the NS300 and held vertically when loading each
sample. Failure to do this can result in incomplete filling of the chamber and imaging problems.
i. Fill a syringe (1 ml to 2.5 ml) with the appropriate sample
Note: large bore syringe bodies (>5 ml) can inadvertently produce high pressures which may damage the top-plate
and should not be used
ii. Remove any air bubbles from the syringe
iii. Hold the laser module vertically, so that the front inlet port is at a lower level than the back outlet port. The
chamber can then be filled slowly against gravity, allowing any bubbles to escape.
iv. Insert the syringe into the front inlet port and introduce the sample slowly into the chamber
Note: When loading the sample cell from dry, liquid should be introduced as slowly as possible, i.e. 1ml in ~10
seconds. This avoids generating pressures which might result in the sample bypassing the seals in the sample
chamber or damaging the top-plate. Loading the sample slowly also limits the introduction of bubbles.
2.2.5.3 Using the syringe-pump
i. Load a syringe with sample and connect it to the luer fitting on the end of the inlet tubing.
ii. Manually load sample into the chamber, following the loading procedure in Section 2.2.5.1 for the flow-cell. For
the O-ring top-plate, the procedures described in Section 2.2.5.2 apply, with the exception that the sample will be
loaded through the inlet tubing rather than via the direct luer fitting on the top-plate.
iii. Once liquid is seen emerging in the waste tubing, place the syringe into the syringe pump holder as described in
the NanoSight Syringe Pump Operating Manual
iv. Operate the syringe pump as described in the manual.

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2.2.6 Mounting the Laser Module
Once the sample is loaded without any air pockets or bubbles, the laser module should be mounted within the main
instrument housing. Rotate the red lever inside the NS300 to the left to allow the laser module to be mounted. Place the
laser module into the slide and gently push forward until it mates with the power connector inside. The red lever inside
should be rotated until vertical to lock the laser module in place, ensuring reproducible positioning.The access door should
then be closed.
a) Laser module mounting / removal b) Laser module locked in position
2.2.7 Loading a New Sample
2.2.7.1 Flow-cell top plate
The flow-cell top-plate has been optimised so that the sample chamber can be cleaned in situ between samples.
i. Rinse the chamber by slowly (i.e. 1ml syringe in 5-10 seconds) loading clean water or a suitable cleaning solution
(please contact Malvern for advice on customised cleaning procedures)
ii. Confirm the cleanliness of the chamber by checking for any particles present in the NTA software image, before
loading the next sample as described in Section 2.2.5.1.
If sample particles persist in the image after rinse through cleaning, or if many particles stuck to the optical flat are visible,
increasing the background noise on the images, the flow-cell should be cleaned manually as described in Section 2.2.4.1.
After samples which may adhere to the optical glass surfaces or between samples which differ significantly in terms of
solvent, particle type or particle loading, it is advisable to dissemble the top plate and rinse and dry thoroughly the optical
element and top plate, to ensure there is no carry-over.
2.2.7.2 O-Ring top plate
When using the O-ring top-plate, the chamber configuration is not optimised for effective flush through cleaning. The top-
plate should always be disassembled and manually cleaned between samples, as described in Section 2.2.4.2, to avoid any
sample carry-over.
Once the sample chamber and any connected tubing have been cleaned, load the next sample as described in Section
2.2.5.2.