Malvern Zetasizer uV User manual

Printed in England MRK1132-01
Malvern Instruments Limited
Enigma Business Park
Grovewood Road, Malvern
Worcs, WR14 1XZ, U.K.
Tel: [phone]
Fax: [phone]
www.malvern.com
user manual
μV
zetasizer
Malvern
user manual
zetasizerzetasizer
μVμV

Zetasizer μV
User Manual
MAN0428 Issue 1.0 December 2008

© Malvern Instruments Ltd. 2003, 2004, 2006, 2007, 2008
Malvern®Instruments makes every effort to ensure that this document is correct. However, due to
Malvern Instruments’ policy of continual product development we are unable to guarantee the
accuracy of this, or any other document after the date of publication. We therefore disclaim all liability
for any changes, errors or omissions after the date of publication. 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
Zetasizer is a registered trademark in the UK and /or other countries, and is owned by
Malvern Instruments Ltd.
Malvern and the green “hills” logo are registered trademarks in the UK and/or other
countries, and is owned by Malvern Instruments Ltd.
Windows 2000 and XP are registered trademarks of the Microsoft Corporation.
Zetasizer, NIBS and M3-PALS are registered trademarks of Malvern Instruments.
Tygon is a registered trademark of Cole Palmer Instruments Company Limited.
Printed in England

Addendum 0076-1.0 Page 1
Addendum to manual
Addendum number ADD0076-1.0
Date 04/02/2009
Manual number MAN0428-1.0
Manual name Zetasizer μV User Manual
This addendum presents new information for taking measurements at ambient
temperatures while using the Flow-mode facility on the Zetasizer μV instrument.
The information below should be inserted as a third paragraph in the Sample -
Temperature section on page 9.16, and as a fifth paragraph in the Flow-mode
SOPs section on page 9.33.
Note
To improve the stability of temperature control when using the system as a
Gel Permeation Chromatography (GPC) detector, the measurement tem-
perature should be set to be 5°C or greater, above or below the ambient
temperature.
Addendum 0076a.fm Page 1 Tuesday, February 24, 2009 4:09 PM

Page 2 Addendum 0076-1.0
Addendum 0076a.fm Page 2 Tuesday, February 24, 2009 4:09 PM

Addendum 0081-1.0 Page 1
Addendum to manual
Addendum number: ADD0081-1.0
Date: 26/05/2010
Manual number: MAN0428-1.0
Manual name: Zetasizer V User Manual
This Addendum presents new information for operating the Zetasizer V instru-
ment. The content should be read in-place of, or in conjunction with, the sections
indicated in the respective manual. The information detailed will be incorporated
into the next manual release for the instrument.
The Zetasizer μV measurement range
The Zetasizer V measurement specifications are described below:
Size range maximum (radius): 0.15nm to 1micron
Molecular weight range: 342Da to 2x107Da
(est from hydrodynamic diameter)
Molecular weight range: 980Da to 2x107Da
(calculated from Debye plot)
For full measurement specifications refer to Appendix 1.
The Zetasizer μV measurement range - Read in place of relevant section on
page 2-1 of the Zetasizer μV User Manual.
ADD0081-1.0 - uV Spec updates.fm Page 1 Wednesday, May 26, 2010 12:01 PM

Page 2 Addendum 0081-1.0
Choosing the correct cell
Caution!
Due to the risk of melting, polystyrene cuvettes must not be used for
measurements above 70°C.
Malvern offers a range of cells for performing measurements with the Zetasizer V.
The choice of cell is dependent upon the type of measurement being performed
and the sample that will be measured. The choices for each measurement type are
outlined below with some discussion on their use.
Size measurements
Choosing the correct cell - Read in place of relevant section on page 4-3 of the
Zetasizer μV User Manual.
Size measurements - Read in place of relevant section on page 4-4 of the
Zetasizer μV User Manual.
Disposable polystyrene
(DTS0012)
Typical solvent Water, Water/ethanol
Optical quality Good to very good
Minimum
Sample volume
1ml
Advantages Low cost
Single use disposable (no
cleaning)
Disadvantages Not resistant to organic
solvents
Unsuitable for use at high
temperatures (above 70°C)
Material Polystyrene
ADD0081-1.0 - uV Spec updates.fm Page 2 Wednesday, May 26, 2010 12:01 PM

Addendum 0081-1.0 Page 3
Specification
Specification - Read in place of relevant section on page A-1 of the Zetasizer
μV User Manual.
Parameter Specification
Size range maximum (radius) * 0.15nm to 1micron
Molecular weight range **
(est from hydrodynamic diameter)
Molecular weight range **
(calculated using Debye plot)
342Da to 2x107Da
980Da to 2x107Da
Minimum sample volume 2µL
Sensitivity at minimum volume 0.1mg/ml of 15kDa protein
High concentration Yes - optically clear samples
SEC detector mode 8µl flow cell
Laser 60mW, 830nm
Temperature range 2°C to 90°C ±0.1 °C
Temperature accuracy 0.1°C at 25°C, 0.2°C at 2°C, 0.5°C at 90°C
Compatible cell temperatures 2°C to 90°C -- Glass and quartz cells
2°C to 70°C -- Plastic disposable cells
Weight 14kg
Dimensions W:D:H, 350mm x 410mm x 170mm
Power 90V-260V AC, 50/60Hz, 100W
Dry air 100kPa (1bar), 4mm o.d. tubing
Computer interface USB
Measurement technique Dynamic light scattering (90 degrees)
Detector Avalanche photodiode, Q.E. >50% at
830nm
Product laser class Class 1 compliant,
IEC 60825-1(1993)+A1(1997)+A2(2001)
Condensation control Purge facility using dry air
Ambient operating conditions
- Humidity
+10 to +35 °C (+50 to 95 °F)
10 to 90% (non-condensing)
* Peak mode range (radius), 0.3nm - 0.5 microns, sample dependent
** Sample and dependent
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Page 4 Addendum 0081-1.0
ADD0081-1.0 - uV Spec updates.fm Page 4 Wednesday, May 26, 2010 12:01 PM

Zetasizer μVPage i
Part 1 - Operator’s Guide
Introduction to this manual
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
How to use this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
Access to the instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Assumed information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Where to get help . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4
What is the Zetasizer μV?
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
What does the Zetasizer μV do? . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
What are Particle Size and Molecular weight? . . . . . . . . . . . . . . . . 2-2
How does the Zetasizer μV work?
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1
How is a Zetasizer measurement performed? . . . . . . . . . . . . . . . . . 3-2
What does the Zetasizer consist of? - Identifying the hardware . . . . 3-5
Navigating the software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-11
A Tutorial - Making a measurement
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1
Quick guide to making a measurement . . . . . . . . . . . . . . . . . . . . . 4-2
Powering up the system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2
Sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Choosing the correct cell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Filling the cell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-7
Inserting the cell. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-9
Making an SOP measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-11
Making a manual measurement . . . . . . . . . . . . . . . . . . . . . . . . . . 4-13
Table of contents

Table of Contents Zetasizer μV
Page ii MAN 0428
The Measurement display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-13
Records and Reports - Viewing the Results
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1
Displaying the results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1
Sample Preparation
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1
Preparing the sample - Size . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1
Preparing the sample - Molecular weight . . . . . . . . . . . . . . . . . . . . 6-2
Count rate meter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4
Part 2 - Supervisor’s Guide
Security
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-1
Initial start-up - set up the administrator. . . . . . . . . . . . . . . . . . . . . 7-2
Enabling security . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-3
User groups. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-4
Users . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-5
Measurement file window - Workspaces
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-1
Measurement file window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-2
Using SOPs
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-1
The SOP Editor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-3
Creating an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-8
Measurement type selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-9
Size SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-10
Molecular weight SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-25
Flow-mode SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-33
Trend SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-35

Zetasizer μVTableofContents
Zetasizer μVPage iii
Extracting an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-37
Modifying an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-38
Distributing an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-38
Advanced features
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-1
Solvent Builder. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-2
SOP Player . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-4
Result averaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-9
Editing the result . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-10
Exporting results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-12
Flow-mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-16
Options dialogue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-21
Expert Advice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-25
Creating Custom Reports
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-1
Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-2
Opening an existing report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-3
Creating a new report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-3
Adding elements to the report . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-4
Setting up the report elements . . . . . . . . . . . . . . . . . . . . . . . . . . 11-5
Selecting elements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-13
Aligning and sizing elements . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-14
Saving a report. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-14
A finished report. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-15
Viewing the new report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-15
Other information shown on a report . . . . . . . . . . . . . . . . . . . . . 11-16
Protecting a report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-16
Protein Utilities
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-1
Debye plot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-2
Molecular weight calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-7
Concentration utilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-10

Table of Contents Zetasizer μV
Page iv MAN 0428
Scattering functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-11
Size theory
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13-1
What is Dynamic Light Scattering? . . . . . . . . . . . . . . . . . . . . . . . 13-1
Operation of the Zetasizer μV - Size measurements . . . . . . . . . . . 13-6
Molecular Weight theory
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14-1
What is Static light scattering? . . . . . . . . . . . . . . . . . . . . . . . . . . 14-1
The Debye plot. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14-4
Part 3 - Appendices
Specification
Specification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-1
Chemical compatibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-2
Unpacking instructions
Installation
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-1
Installing the Zetasizer μV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-2
Changing the computer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-2
Regulatory Statements
Disposal of Electrical & Electronic Equipment . . . . . . . . . . . . . . . . . D-1
FCC Notice (US only) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . D-2
Canadian Regulatory Information. . . . . . . . . . . . . . . . . . . . . . . . . . D-3
VCCI acceptance (Japan only). . . . . . . . . . . . . . . . . . . . . . . . . . . . D-3

Part 1 -
Operator’s Guide

Zetasizer μVPage 1-1
1
Introduction
to this manual
Introduction
This manual covers the operation and maintenance of the Zetasizer μVparticle
analyser.
Note
The Zetasizer model, Serial number, software and firmware version can be
found by left-clicking the μV icon in the right corner of the status bar.
The aim of this manual is to:
Identify what the instrument is.
Explain in simple terms how it works.
Explain how the instrument should be used to make a measurement.
Identify the user maintenance procedures.
How to use this manual
Read this manual together with the Essentials manual, which describes Site
Requirements, Health and Safety and Maintenance information.
Warning!
The instrument and the samples to be measured may be hazardous if mis-
used. Users must read the Health and Safety information in the Essen-
tials manual before operating the system.

Chapter 1 Introduction to this manual
Page 1-2 MAN 0428
1
We recommend reading both these manuals fully before starting the first measure-
ment, though if more familiar with particle size analysers, jump straight to Chap-
ter 4. This chapter gives practical details on making measurements.
This manual is divided into three sections as follows.
Part 1 – Operator’s guide
This guide and the Essentials manual contain all the information required for the
operator to use the Zetasizer μV instrument.
Topics covered are: What is the Zetasizer μV instrument, what are the components
of the Zetasizer μV, what it does, instructions on using the instrument and the soft-
ware, basic measurement and maintenance procedures.
Part 2 – Supervisor’s guide
The Supervisor’s guide concentrates on the administration of the Zetasizer μV and
covers some of the more advanced features. It also provides a greater insight into
the measurement procedures and expands on the analysis theories.
Topics covered are: security aspects, use of Standard Operating Procedures
(SOPs), organising measurement files and discussion on the analysis theories used
– Size and Molecular weight.
It is recommended that the supervisor should also read Part 1 – Operator’s guide.
Part 3 - Appendices
This contains supplementary information not necessary for the general operation
of the system.
More detail on the Zetasizer software can be found by using the on-line Help
within the software.

Introduction to this manual Chapter 1
Zetasizer μVPage 1-3
Access to the instrument
Within this manual, reference is made to the various people who will have access to
the instrument.
Malvern personnel
Malvern personnel (service engineers, representatives, etc.) have full access to the
instrument and are the only people authorised to perform all service procedures
that may require the removal of the covers.
Warning!
Removal of the main covers by unauthorised personnel, even a supervisor,
will invalidate the warranty of the instrument.
Supervisor
The supervisor is responsible for the management and safety of the instrument and
its operation. The supervisor also trains the operators. They can perform all user
maintenance routines identified in the Essentials manual.
Under no circumstances, should the supervisor remove the main cover of the
instrument.
Operator
An operator is a person trained in the use of the system. The operator can perform
all user maintenance routines identified in the Essentials manual, except chang-
ing the fuse.
Under no circumstances, should the operator remove the main cover of the instru-
ment.
Warning!
Failure to follow these guidelines could result in exposure to hazardous
voltages and laser radiation.
Assumed information
Naming convention
The Zetasizer μV will either be referred to in full, or as “the instrument”.

Chapter 1 Introduction to this manual
Page 1-4 MAN 0428
The combination of the Zetasizer μV instrument, the computer and Zetasizer soft-
ware may be referred to as the “the system”.
Cells and Cuvettes
Any device for holding and measuring the sample in the instrument will generally
be referred to as a “cell”. This includes dip cells and all kinds of cuvettes used (i.e.
glass, small volume, disposable) unless a proper description is more appropriate.
Solvents and dispersants
To reflect the intended protein analysis applications of the system, the term
“solvent” is used throughout this manual to refer to any fluid used to disperse a
sample’s particles for measurement. However, solvent can also be taken to mean
“dispersant” where this is more applicable to the sample type.
Menu commands
Menu commands from the Zetasizer software are referred to in the form main
menu-menu item. As an example, the command Tools-Options refers to select-
ing the Options item in the Tools menu. Menu commands are shown in bold
text.
Where to get help
Manuals and Help files
The primary sources of help for the Zetasizer μV are the Essentials manual, this
manual and the on-line help system within the software. This manual is designed
to give an overview of the system as a whole, while the on-line help system is
designed to give detailed information on the Zetasizer software.
Each dialogue within the Zetasizer software has a Help button that gives informa-
tion specific to that dialogue.
Essentials Manual
This manual covers:
Site requirements – all the physical requirements for positioning the system.
Information is given on; services requirements (air, water, number of power
sockets, etc.), environmental requirements (temperature, humidity etc.) and
physical requirements (space requirements etc.).
Health and safety – this must be read by all users of the system. It details all
safety issues for the optical unit and all accessories.