Particle size analysis -- Image analysis methods

ISO 13322-1:2014 is applicable to the analysis of images for the purpose of determining particle size distributions where the velocity of the particles against the axis of the optical system of the imaging device is zero. The particles are appropriately dispersed and fixed in the object plane of the instrument. The field of view may sample the object plane dynamically either by moving the sample support or the camera provided this can be accomplished without any motion effects on the image. Captured images can be analysed subsequently. ISO 13322-1:2014 concentrates upon the analysis of digital images created from either light or electron detection systems. It considers only image evaluation methods using complete pixel counts.

Analyse granulométrique -- Méthodes par analyse d'images

Granulometrijska analiza - Metode analize slike - 1. del: Statične metode analize slike

Ta del standarda ISO 13322 velja za analize slik za določanje razporeditve velikosti delcev, kadar je hitrost delcev glede na os optičnega sistema naprave za slikanje enaka nič. Delci so ustrezno razpršeni in fiksni v predmetni ravnini instrumenta. Vidno polje lahko vzorči predmetno ravnino dinamično s premikanjem podpore vzorca ali predložene kamere. To je mogoče doseči brez kakršnih koli učinkov gibanja na sliko. Posnete slike je mogoče analizirati pozneje.
Ta del standarda ISO 13322 se osredotoča na analizo digitalnih slik, ustvarjenih s sistemi za zaznavanje svetlobe ali elektronov. Ne zajema metode ustvarjanja slike, čeprav je nastavitev zaznavanja, ki je izbrana skupaj s kalibracijo, pomembna za natančnost določanja velikosti delcev. Ta del standarda ISO 13322 zajema samo metode ocenjevanja slik z uporabo popolnega štetja pikslov. Vrsta porazdelitve (glede na število ali prostornino) skupaj s širino porazdelitve velikosti delcev ima bistven vpliv na število delcev za merjenje, da se zagotovi natančnost v okviru določenih mejah zaupanja. Primer je prikazan v prilogi A. Avtomatizacija analize je mogoča, da se izmeri zadostno število delcev za zahtevano stopnjo natančnosti.
Ta del standarda ISO 13322 ne obravnava priprave vzorca. Kljub temu so podvzorčenje, razpršitev in predstavitev delcev za merjenje ključni deli operativne verige dejanj, ki so potrebna za zagotovitev pravilnosti in natančnosti končnih rezultatov.

General Information

Status
Published
Publication Date
11-May-2014
Current Stage
6060 - International Standard published
Start Date
10-Apr-2014
Completion Date
12-May-2014

RELATIONS

Buy Standard

Standard
ISO 13322-1:2014 - Particle size analysis -- Image analysis methods
English language
24 pages
sale 15% off
Preview
sale 15% off
Preview
Standard
ISO 13322-1:2015 - natisnjeno za čitalnico
English language
29 pages
sale 10% off
Preview
sale 10% off
Preview

e-Library read for
1 day

Standards Content (sample)

INTERNATIONAL ISO
STANDARD 13322-1
Second edition
2014-05-15
Particle size analysis — Image analysis
methods —
Part 1:
Static image analysis methods
Analyse granulométrique — Méthodes par analyse d’images —
Partie 1: Méthodes par analyse d’images statiques
Reference number
ISO 13322-1:2014(E)
ISO 2014
---------------------- Page: 1 ----------------------
ISO 13322-1:2014(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2014

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2014 – All rights reserved
---------------------- Page: 2 ----------------------
ISO 13322-1:2014(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions and list of symbols ................................................................................................................................. 1

3.1 Terms and definitions ....................................................................................................................................................................... 1

3.2 Symbols ......................................................................................................................................................................................................... 4

4 Preparation for image capture .............................................................................................................................................................. 5

4.1 Introduction .............................................................................................................................................................................................. 5

4.2 Procedures .................................................................................................................................................................................................. 5

5 Sample preparation demands for method description ................................................................................................ 6

5.1 Sample splitting and reduction ................................................................................................................................................. 6

5.2 Touching particles ................................................................................................................................................................................ 6

5.3 Particle distribution............................................................................................................................................................................ 6

5.4 Number of particles to be counted ........................................................................................................................................ 6

5.5 Particle suspending fluid ................................................................................................................................................................ 7

6 Quality of captured images ........................................................................................................................................................................ 7

6.1 General ........................................................................................................................................................................................................... 7

6.2 Pixels per particle ................................................................................................................................................................................. 7

7 Image analysis ......................................................................................................................................................................................................... 8

7.1 General ........................................................................................................................................................................................................... 8

7.2 Size classes and magnification .................................................................................................................................................. 8

8 Counting procedure ........................................................................................................................................................................................... 9

8.1 General ........................................................................................................................................................................................................... 9

8.2 Particle image edges ........................................................................................................................................................................... 9

8.3 Particles cut by the edge of the measurement frame..........................................................................................10

8.4 Touching particles .............................................................................................................................................................................11

8.5 Measurements ......................................................................................................................................................................................12

9 Calculation of the particle size results ........................................................................................................................................12

10 Calibration and traceability ...................................................................................................................................................................12

10.1 General ........................................................................................................................................................................................................12

10.2 Recommendations and requirements ..............................................................................................................................13

11 Accuracy .....................................................................................................................................................................................................................14

11.1 General ........................................................................................................................................................................................................14

11.2 Reference materials .........................................................................................................................................................................14

11.3 Instrument preparation ...............................................................................................................................................................14

11.4 Qualification test ................................................................................................................................................................................15

11.5 Qualification acceptance ..............................................................................................................................................................15

12 Test report ................................................................................................................................................................................................................15

Annex A (informative) Estimation of the number of particles to be counted for a given accuracy ..17

Annex B (informative) Common segmentation methods for particle edge detection....................................22

Annex C (informative) Flow chart showing a typical image analysis method.........................................................23

Bibliography .............................................................................................................................................................................................................................24

© ISO 2014 – All rights reserved iii
---------------------- Page: 3 ----------------------
ISO 13322-1:2014(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any

patent rights identified during the development of the document will be in the Introduction and/or on

the ISO list of patent declarations received (see www.iso.org/patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation on the meaning of ISO specific terms and expressions related to conformity

assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers

to Trade (TBT) see the following URL: Foreword - Supplementary information

The committee responsible for this document is ISO/TC 24, Particle characterization including sieving,

Subcommittee SC 4, Particle characterization.

This second edition cancels and replaces the first edition (ISO 13322-1:2004), which has been technically

revised.

ISO 13322 consists of the following parts, under the general title Particle size analysis — Image analysis

methods:
— Part 1: Static image analysis methods
— Part 2: Dynamic image analysis methods
iv © ISO 2014 – All rights reserved
---------------------- Page: 4 ----------------------
ISO 13322-1:2014(E)
Introduction

The purpose of this part of ISO 13322 is to give guidance when using images for particle size analysis.

Image analysis is a technique that has gained popularity in different applications. The aim of this part

of ISO 13322 is to give a standardized description of the technique used and its validation. This part of

ISO 13322 does not describe specific instruments and is restricted to those parts of the acquisition of

images that are relevant to the accuracy of the particle size analysis.

This part of ISO 13322 includes methods of calibration verification and recommends using a certified

standard as a reference scale. However it is sensible to make some measurements on particles under

study, or other reference objects, of known size so that the likely systematic uncertainties introduced by

the equipment can be assessed.

Errors introduced at all stages of the analysis from sub-division of the sample to generation of the final

result add to the total uncertainty of measurements and it is important to obtain estimates for the

uncertainty arising from each stage.

Essential operations are identified to ensure that measurements made conform to this part of ISO 13322

and are traceable.
© ISO 2014 – All rights reserved v
---------------------- Page: 5 ----------------------
INTERNATIONAL STANDARD ISO 13322-1:2014(E)
Particle size analysis — Image analysis methods —
Part 1:
Static image analysis methods
1 Scope

This part of ISO 13322 is applicable to the analysis of images for the purpose of determining particle

size distributions where the velocity of the particles against the axis of the optical system of the imaging

device is zero. The particles are appropriately dispersed and fixed in the object plane of the instrument.

The field of view may sample the object plane dynamically either by moving the sample support or the

camera provided this can be accomplished without any motion effects on the image. Captured images

can be analysed subsequently.

This part of ISO 13322 concentrates upon the analysis of digital images created from either light or

electron detection systems. It does not address the method of creating the image although the detection

settings chosen together with its calibration are important to particle sizing accuracy. This part of

ISO 13322 considers only image evaluation methods using complete pixel counts.

Both the type of distribution, (by number or by volume) together with the width of the particle size

distribution has a very material influence upon the number of particles to be measured to secure the

desired accuracy within the specified confidence limits. An example is shown in Annex A.

Automation of the analysis is possible in order to measure sufficient particle numbers for a required

degree of precision.

This part of ISO 13322 does not address the sample preparation. However, the sub sampling, dispersion

and presentation of particles to be measured are a vital part of the operational chain of actions necessary

to ensure accuracy and precision of any final result.

NOTE Further details about sampling and sample preparation can be found in ISO 14887 and ISO 14488.

2 Normative references

The following documents, in whole or in part, are normatively referenced in this document and are

indispensable for its application. For dated references, only the edition cited applies. For undated

references, the latest edition of the referenced document (including any amendments) applies.

ISO 9276-1, Representation of results of particle size analysis — Part 1: Graphical representation

ISO 9276-2, Representation of results of particle size analysis — Part 2: The calculations of average particle

sizes/diameters and moments from particle size distributions

ISO 14488, Particulate materials — Sampling and sample splitting for the determination of particulate

properties
3 Terms and definitions and list of symbols
3.1 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
© ISO 2014 – All rights reserved 1
---------------------- Page: 6 ----------------------
ISO 13322-1:2014(E)
3.1.1
area equivalent diameter
diameter of a circle having the same area as the projected image of the particle

Note 1 to entry: It is also known as the Heywood diameter or as the equivalent circular diameter.

3.1.2
binary image

digitized image consisting of an array of pixels, each of which has a value of 0 or 1, whose values are

normally represented by dark and bright regions on the display screen or by the use of two distinct

colours
3.1.3
contrast (of an image)

difference between the intensity of the particle image with respect to the

background near to the particle
3.1.4
edge detection
methods used to detect transition between objects and background
Note 1 to entry: See segmentation method (3.1.13).
3.1.5
Feret diameter

distance between two parallel tangents on opposite sides of the image of a particle

3.1.6
field of view
field which is viewed by the viewing device

Note 1 to entry: The full image frame of a digital imaging device corresponds to its field of view.

SEE: Figure 1.
3.1.7
grey image
image in which multiple grey level values are permitted for each pixel
3.1.8
image analysis

processing and data reduction operation which yields a numerical or logical result from an image

3.1.9
measurement field
field which is composed by the set of all measurement frames
SEE: Figure 1.
3.1.10
measurement frame

selected area from the field of view in which particles are sized and counted for image analysis

SEE: Figure 1.
3.1.11
pixel
picture element

individual sample in a digital image that has been formed by uniform sampling in both the horizontal

and vertical directions
2 © ISO 2014 – All rights reserved
---------------------- Page: 7 ----------------------
ISO 13322-1:2014(E)
3.1.12
raster pattern
scanning order of measurement frames in the total measurement field
SEE: Figure 1.
3.1.13
segmentation method
strategy employed to separate the objects of interest from their surroundings
Note 1 to entry: Method of dividing the particle image from the background.
Note 2 to entry: See edge detection (3.1.4).
3.1.14
threshold

grey level value which is set to discriminate objects of interest from background

Key
1 measurement frame
2 field of view
3 raster pattern of measurement frames
4 measurement field
X enlarged view of a field of view

Figure 1 — Relationship between the terms “field of view”, “measurement frame”, “raster

pattern” and “measurement field”
© ISO 2014 – All rights reserved 3
---------------------- Page: 8 ----------------------
ISO 13322-1:2014(E)
3.2 Symbols
A projected area of particle i
α horizontal calibration factor
α vertical calibration factor
d minimum feature length
d diameter of a circle
N number of particles to be measured
n measured number of pixels within a circle
n numbers of particles in size interval Δx
j j

P probability that particle i exists in the measuring frame (also called Miles-Lantuéjoul factor)

φ shape descriptor
σ standard deviation
V volume of particle i
x area equivalent diameter of particle i
A,i
x horizontal Feret diameter of object
x vertical Feret diameter of object
x dimension of particle i
x longest dimension of particle i, also called maximum Feret diameter
Fmax,i
x shortest dimension of particle i, also called minimum Feret diameter
Fmin,i
x horizontal dimension of object
x horizontal dimension of object in SI unit
1,m
x horizontal dimension of object in pixel
1,p
x vertical dimension of object
x vertical dimension of object in SI unit
2,m
x vertical dimension of object in pixel
2,p

x particle size corresponding to 10 % of the cumulative undersize distribution by volume

10,3

x particle size corresponding to 90 % of the cumulative undersize distribution by volume

90,3
Z horizontal side length of the rectangular measurement frame
Z vertical side length of the rectangular measurement frame
4 © ISO 2014 – All rights reserved
---------------------- Page: 9 ----------------------
ISO 13322-1:2014(E)
4 Preparation for image capture
4.1 Introduction

A pre-requisite for accurate particle size measurement using this method requires a full understanding

of the settings and calibration applied within the image capture device as well as a consideration of the

purpose for conducting the measurement.

The final settings and calibration of the image capture device need to be established via an iterative

approach. The size range of the particles within an unknown test sample has an influence upon the

settings required within the image capturing device. These remain unknown until the first image has

been taken, the result observed and the necessary adjustments to the image capture device to achieve

the desired accuracy of particle size measurement required. A fully trained operator shall conduct the

assessment.

The imaging instrument should be set up and operated in accordance with the manufacturer’s

recommendations considering the conditions prevailing.

In order to achieve accurate particle size measurements it is preferred that the illumination be uniform

over the total field of view and of a type designed to create images of high contrast. The magnification

should be such as to provide a minimum number of pixels for the smallest particle consistent with the

accuracy demanded and set to achieve a sharp focus. The number of pixels for the smallest dimension of

a particle is relevant for cases where linear dimensions or combinations thereof are measured.

Distortion in the image might arise from a number of causes, but its presence and effect on the image

may be determined by selecting known sized particles or other reference objects of similar optical

properties at a number of points and orientations in the field of view. It is important to note that the

measurements made provide only two-dimensional, X and Y, information.
4.2 Procedures

The operator should decide why the result of the image analysis is required. Is a size distribution

by the number of particles in each size class required or is the volume of particles in each size class

the requirement? What accuracy and precision is required for the final result? These decisions will

have a significant influence upon the choice of settings and the method employed in conducting the

measurement.

For each material to be analysed and for each instrument employed the person conducting the analysis

shall ensure that the following procedures are followed.

a) Ensure an adequate calibration for both the X and Y-axis of the measurement frame exists for the

imaging instrument being employed, preferably by using a certified graticule or equivalent reference

of equal standing.

b) Ensure that the optical magnification employed is suitable and such as the image of the smallest

particle to be measured covers a sufficient number of pixels to support the required accuracy of

measurement.

c) Ensure that the illumination method and setting of any focus is correctly established to give a good

contrast and uniformity of illumination of any image gathered.

d) Ensure that the number of particles within the measurement frame is such as to minimize the

number of touching particles.

e) Ensure that a sufficient number of images of separate aliquot samples are gathered to provide a

suitable total number of particles with respect to the type of distribution, number or volume based,

and the width of the particle size distribution (see ISO 14488) and that they contain an adequate

statistical number of the largest particle of the target material (see Annex A).
© ISO 2014 – All rights reserved 5
---------------------- Page: 10 ----------------------
ISO 13322-1:2014(E)

f) Some implementations of the image analysis technique employ a large area X, Y servo or manually

controlled sample slide assembly. Such large slides enable many measurement frames of the particles

deposited to be examined. Should the method of fully separate measurement frames be employed

then any frame overlap shall be avoided. If the method of overlapping measurement frames, or

other methods of analysing particles that interact with the measurement frame edge is used, then

procedures shall be employed to ensure that each particle is only included into the total count for

the appropriate size class, once. For more information, see 8.3.

g) If appropriate, ensure that the image quality consisting of illumination, focus and magnification has

not changed at the end of the measurement. The requirement for this step depends on the variability

of the instrument employed.

h) For the case when image analysis is to be used for certified reference material measurement at the

end of the image gathering procedure, the calibration outlined in a) should be repeated and any

measured deviation recorded.

i) All the conditions, set up or established, for the target material shall be fully documented.

5 Sample preparation demands for method description
5.1 Sample splitting and reduction

As only a small amount of material is needed to prepare a test sample, this should be sub-divided from

the whole sample in a manner that ensures that the test sample is representative of the whole as specified

in ISO 14488.
5.2 Touching particles

In order to assess the degree of touching particles, a suitable optical resolution setting of the imaging

system should be chosen. The optical resolution should also meet the criteria set out in 4.2 b).

The number of particles touching each other should be minimized. It is a prime requirement of the

method that measurements shall be made on isolated particles. Touching particles measured as one

particle without a proper separation will introduce error.

It is often not possible to reliably detect touching particles by image analysis alone, but the influence

of touching particles on the result can be investigated experimentally by increasing or decreasing the

number of particles per image. If the number of particles cannot be changed, the influence on the results

can be investigated using a reference material with similar size and shape.
5.3 Particle distribution

There should be an adequate distribution of particles in the field of view. It may be necessary to examine

several fields of view if a large total particle count is required. The whole area of the measurement

field should be examined to ascertain whether there is noticeable segregation of particles (by size). The

requirements set out in 4.2 f) should be followed.
5.4 Number of particles to be counted

The number of particles to be analysed depends upon whether the final result is a particle size

distribution by the number or by the volume of particles. Considerable care has to be exercised in order

to ensure that the analysis is representative of the bulk sample as described in ISO 14488. This can be

demonstrated by splitting the bulk sample into at least three test samples. Each test sample should

contain sufficient particle numbers for a full measurement. Statistical analysis of the data will reveal the

repeatability of the method including sampling and dispersion.
NOTE See Annex A for more information.
6 © ISO 2014 – All rights reserved
---------------------- Page: 11 ----------------------
ISO 13322-1:2014(E)
5.5 Particle suspending fluid

It is likely that a large number of particle measurements will be of particles presented in a gas where an

adequate image contrast should be ensured. Should particle presentation require a liquid suspension

then it is preferred that such liquids be clean, particle-free, transparent and have a refractive index

as different as possible from the refractive index of particles to enhance the image contrast. Particles

presented in a mixed optical background such as in biological specimens, may require dynamic particle

by particle threshold selection.

WARNING — Automated particle by particle parameter selection applied for segmentation as

envisaged when mixed optical backgrounds are required cannot be validated for true particle

size and may result in particle size bias and reduced accuracy.

CAUTION — Particle systems of mixed optical properties may have inaccurate particle sizes

attributed to them due to possible threshold setting errors introducing particle size bias.

6 Quality of captured images
6.1 General

It is important for the analysis that the particles in the captured images are well dispersed. The number

of overlapping or touching particles reduces with the concentration of the particles in the frame (see

5.2). This is in conflict with the requirements of having a large particle number to obtain a high degree

of precision. A compromise should be established.

The contrast achieved should be consistent with the level of accuracy required. The difference between

the brightness of the particle and its background shall be a few times of the resolution of the grey level

image from zero to maximum brightness signal.

The accuracy of the results is strongly affected by the number of pixels for each particle image. When only

the projected area of the particles is measured as an average value, a few pixels may provide acceptable

results for the smallest particles. Higher pixel numbers are required for accurate information about

each individual particle.
6.2 Pixels per particle

Both the number of pixels forming the image of the particle and the relative position of the centring of

the image with respect to the fixed pixel pattern can have a material influence upon the final particle

size assessed from each particle image.

Image analysis can be a method of choice for the certification of reference materials. It also can be the

method of choice for general measurements. The conditions required to achieve a defined

...

SLOVENSKI STANDARD
SIST ISO 13322-1:2015
01-februar-2015
1DGRPHãþD
SIST ISO 13322-1:2006

*UDQXORPHWULMVNDDQDOL]D0HWRGHDQDOL]HVOLNHGHO6WDWLþQHPHWRGHDQDOL]H

VOLNH

Particle size analysis - Image analysis methods - Part 1: Static image analysis methods

Analyse granulométrique - Méthodes par analyse d'images - Partie 1: Méthodes par
analyse d'images statiques
Ta slovenski standard je istoveten z: ISO 13322-1:2014
ICS:
19.120 Analiza velikosti delcev. Particle size analysis. Sieving
Sejanje
SIST ISO 13322-1:2015 en

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST ISO 13322-1:2015
---------------------- Page: 2 ----------------------
SIST ISO 13322-1:2015
INTERNATIONAL ISO
STANDARD 13322-1
Second edition
2014-05-15
Particle size analysis — Image analysis
methods —
Part 1:
Static image analysis methods
Analyse granulométrique — Méthodes par analyse d’images —
Partie 1: Méthodes par analyse d’images statiques
Reference number
ISO 13322-1:2014(E)
ISO 2014
---------------------- Page: 3 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2014

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2014 – All rights reserved
---------------------- Page: 4 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions and list of symbols ................................................................................................................................. 1

3.1 Terms and definitions ....................................................................................................................................................................... 1

3.2 Symbols ......................................................................................................................................................................................................... 4

4 Preparation for image capture .............................................................................................................................................................. 5

4.1 Introduction .............................................................................................................................................................................................. 5

4.2 Procedures .................................................................................................................................................................................................. 5

5 Sample preparation demands for method description ................................................................................................ 6

5.1 Sample splitting and reduction ................................................................................................................................................. 6

5.2 Touching particles ................................................................................................................................................................................ 6

5.3 Particle distribution............................................................................................................................................................................ 6

5.4 Number of particles to be counted ........................................................................................................................................ 6

5.5 Particle suspending fluid ................................................................................................................................................................ 7

6 Quality of captured images ........................................................................................................................................................................ 7

6.1 General ........................................................................................................................................................................................................... 7

6.2 Pixels per particle ................................................................................................................................................................................. 7

7 Image analysis ......................................................................................................................................................................................................... 8

7.1 General ........................................................................................................................................................................................................... 8

7.2 Size classes and magnification .................................................................................................................................................. 8

8 Counting procedure ........................................................................................................................................................................................... 9

8.1 General ........................................................................................................................................................................................................... 9

8.2 Particle image edges ........................................................................................................................................................................... 9

8.3 Particles cut by the edge of the measurement frame..........................................................................................10

8.4 Touching particles .............................................................................................................................................................................11

8.5 Measurements ......................................................................................................................................................................................12

9 Calculation of the particle size results ........................................................................................................................................12

10 Calibration and traceability ...................................................................................................................................................................12

10.1 General ........................................................................................................................................................................................................12

10.2 Recommendations and requirements ..............................................................................................................................13

11 Accuracy .....................................................................................................................................................................................................................14

11.1 General ........................................................................................................................................................................................................14

11.2 Reference materials .........................................................................................................................................................................14

11.3 Instrument preparation ...............................................................................................................................................................14

11.4 Qualification test ................................................................................................................................................................................15

11.5 Qualification acceptance ..............................................................................................................................................................15

12 Test report ................................................................................................................................................................................................................15

Annex A (informative) Estimation of the number of particles to be counted for a given accuracy ..17

Annex B (informative) Common segmentation methods for particle edge detection....................................22

Annex C (informative) Flow chart showing a typical image analysis method.........................................................23

Bibliography .............................................................................................................................................................................................................................24

© ISO 2014 – All rights reserved iii
---------------------- Page: 5 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any

patent rights identified during the development of the document will be in the Introduction and/or on

the ISO list of patent declarations received (see www.iso.org/patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation on the meaning of ISO specific terms and expressions related to conformity

assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers

to Trade (TBT) see the following URL: Foreword - Supplementary information

The committee responsible for this document is ISO/TC 24, Particle characterization including sieving,

Subcommittee SC 4, Particle characterization.

This second edition cancels and replaces the first edition (ISO 13322-1:2004), which has been technically

revised.

ISO 13322 consists of the following parts, under the general title Particle size analysis — Image analysis

methods:
— Part 1: Static image analysis methods
— Part 2: Dynamic image analysis methods
iv © ISO 2014 – All rights reserved
---------------------- Page: 6 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
Introduction

The purpose of this part of ISO 13322 is to give guidance when using images for particle size analysis.

Image analysis is a technique that has gained popularity in different applications. The aim of this part

of ISO 13322 is to give a standardized description of the technique used and its validation. This part of

ISO 13322 does not describe specific instruments and is restricted to those parts of the acquisition of

images that are relevant to the accuracy of the particle size analysis.

This part of ISO 13322 includes methods of calibration verification and recommends using a certified

standard as a reference scale. However it is sensible to make some measurements on particles under

study, or other reference objects, of known size so that the likely systematic uncertainties introduced by

the equipment can be assessed.

Errors introduced at all stages of the analysis from sub-division of the sample to generation of the final

result add to the total uncertainty of measurements and it is important to obtain estimates for the

uncertainty arising from each stage.

Essential operations are identified to ensure that measurements made conform to this part of ISO 13322

and are traceable.
© ISO 2014 – All rights reserved v
---------------------- Page: 7 ----------------------
SIST ISO 13322-1:2015
---------------------- Page: 8 ----------------------
SIST ISO 13322-1:2015
INTERNATIONAL STANDARD ISO 13322-1:2014(E)
Particle size analysis — Image analysis methods —
Part 1:
Static image analysis methods
1 Scope

This part of ISO 13322 is applicable to the analysis of images for the purpose of determining particle

size distributions where the velocity of the particles against the axis of the optical system of the imaging

device is zero. The particles are appropriately dispersed and fixed in the object plane of the instrument.

The field of view may sample the object plane dynamically either by moving the sample support or the

camera provided this can be accomplished without any motion effects on the image. Captured images

can be analysed subsequently.

This part of ISO 13322 concentrates upon the analysis of digital images created from either light or

electron detection systems. It does not address the method of creating the image although the detection

settings chosen together with its calibration are important to particle sizing accuracy. This part of

ISO 13322 considers only image evaluation methods using complete pixel counts.

Both the type of distribution, (by number or by volume) together with the width of the particle size

distribution has a very material influence upon the number of particles to be measured to secure the

desired accuracy within the specified confidence limits. An example is shown in Annex A.

Automation of the analysis is possible in order to measure sufficient particle numbers for a required

degree of precision.

This part of ISO 13322 does not address the sample preparation. However, the sub sampling, dispersion

and presentation of particles to be measured are a vital part of the operational chain of actions necessary

to ensure accuracy and precision of any final result.

NOTE Further details about sampling and sample preparation can be found in ISO 14887 and ISO 14488.

2 Normative references

The following documents, in whole or in part, are normatively referenced in this document and are

indispensable for its application. For dated references, only the edition cited applies. For undated

references, the latest edition of the referenced document (including any amendments) applies.

ISO 9276-1, Representation of results of particle size analysis — Part 1: Graphical representation

ISO 9276-2, Representation of results of particle size analysis — Part 2: The calculations of average particle

sizes/diameters and moments from particle size distributions

ISO 14488, Particulate materials — Sampling and sample splitting for the determination of particulate

properties
3 Terms and definitions and list of symbols
3.1 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
© ISO 2014 – All rights reserved 1
---------------------- Page: 9 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
3.1.1
area equivalent diameter
diameter of a circle having the same area as the projected image of the particle

Note 1 to entry: It is also known as the Heywood diameter or as the equivalent circular diameter.

3.1.2
binary image

digitized image consisting of an array of pixels, each of which has a value of 0 or 1, whose values are

normally represented by dark and bright regions on the display screen or by the use of two distinct

colours
3.1.3
contrast (of an image)

difference between the intensity of the particle image with respect to the

background near to the particle
3.1.4
edge detection
methods used to detect transition between objects and background
Note 1 to entry: See segmentation method (3.1.13).
3.1.5
Feret diameter

distance between two parallel tangents on opposite sides of the image of a particle

3.1.6
field of view
field which is viewed by the viewing device

Note 1 to entry: The full image frame of a digital imaging device corresponds to its field of view.

SEE: Figure 1.
3.1.7
grey image
image in which multiple grey level values are permitted for each pixel
3.1.8
image analysis

processing and data reduction operation which yields a numerical or logical result from an image

3.1.9
measurement field
field which is composed by the set of all measurement frames
SEE: Figure 1.
3.1.10
measurement frame

selected area from the field of view in which particles are sized and counted for image analysis

SEE: Figure 1.
3.1.11
pixel
picture element

individual sample in a digital image that has been formed by uniform sampling in both the horizontal

and vertical directions
2 © ISO 2014 – All rights reserved
---------------------- Page: 10 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
3.1.12
raster pattern
scanning order of measurement frames in the total measurement field
SEE: Figure 1.
3.1.13
segmentation method
strategy employed to separate the objects of interest from their surroundings
Note 1 to entry: Method of dividing the particle image from the background.
Note 2 to entry: See edge detection (3.1.4).
3.1.14
threshold

grey level value which is set to discriminate objects of interest from background

Key
1 measurement frame
2 field of view
3 raster pattern of measurement frames
4 measurement field
X enlarged view of a field of view

Figure 1 — Relationship between the terms “field of view”, “measurement frame”, “raster

pattern” and “measurement field”
© ISO 2014 – All rights reserved 3
---------------------- Page: 11 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
3.2 Symbols
A projected area of particle i
α horizontal calibration factor
α vertical calibration factor
d minimum feature length
d diameter of a circle
N number of particles to be measured
n measured number of pixels within a circle
n numbers of particles in size interval Δx
j j

P probability that particle i exists in the measuring frame (also called Miles-Lantuéjoul factor)

φ shape descriptor
σ standard deviation
V volume of particle i
x area equivalent diameter of particle i
A,i
x horizontal Feret diameter of object
x vertical Feret diameter of object
x dimension of particle i
x longest dimension of particle i, also called maximum Feret diameter
Fmax,i
x shortest dimension of particle i, also called minimum Feret diameter
Fmin,i
x horizontal dimension of object
x horizontal dimension of object in SI unit
1,m
x horizontal dimension of object in pixel
1,p
x vertical dimension of object
x vertical dimension of object in SI unit
2,m
x vertical dimension of object in pixel
2,p

x particle size corresponding to 10 % of the cumulative undersize distribution by volume

10,3

x particle size corresponding to 90 % of the cumulative undersize distribution by volume

90,3
Z horizontal side length of the rectangular measurement frame
Z vertical side length of the rectangular measurement frame
4 © ISO 2014 – All rights reserved
---------------------- Page: 12 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
4 Preparation for image capture
4.1 Introduction

A pre-requisite for accurate particle size measurement using this method requires a full understanding

of the settings and calibration applied within the image capture device as well as a consideration of the

purpose for conducting the measurement.

The final settings and calibration of the image capture device need to be established via an iterative

approach. The size range of the particles within an unknown test sample has an influence upon the

settings required within the image capturing device. These remain unknown until the first image has

been taken, the result observed and the necessary adjustments to the image capture device to achieve

the desired accuracy of particle size measurement required. A fully trained operator shall conduct the

assessment.

The imaging instrument should be set up and operated in accordance with the manufacturer’s

recommendations considering the conditions prevailing.

In order to achieve accurate particle size measurements it is preferred that the illumination be uniform

over the total field of view and of a type designed to create images of high contrast. The magnification

should be such as to provide a minimum number of pixels for the smallest particle consistent with the

accuracy demanded and set to achieve a sharp focus. The number of pixels for the smallest dimension of

a particle is relevant for cases where linear dimensions or combinations thereof are measured.

Distortion in the image might arise from a number of causes, but its presence and effect on the image

may be determined by selecting known sized particles or other reference objects of similar optical

properties at a number of points and orientations in the field of view. It is important to note that the

measurements made provide only two-dimensional, X and Y, information.
4.2 Procedures

The operator should decide why the result of the image analysis is required. Is a size distribution

by the number of particles in each size class required or is the volume of particles in each size class

the requirement? What accuracy and precision is required for the final result? These decisions will

have a significant influence upon the choice of settings and the method employed in conducting the

measurement.

For each material to be analysed and for each instrument employed the person conducting the analysis

shall ensure that the following procedures are followed.

a) Ensure an adequate calibration for both the X and Y-axis of the measurement frame exists for the

imaging instrument being employed, preferably by using a certified graticule or equivalent reference

of equal standing.

b) Ensure that the optical magnification employed is suitable and such as the image of the smallest

particle to be measured covers a sufficient number of pixels to support the required accuracy of

measurement.

c) Ensure that the illumination method and setting of any focus is correctly established to give a good

contrast and uniformity of illumination of any image gathered.

d) Ensure that the number of particles within the measurement frame is such as to minimize the

number of touching particles.

e) Ensure that a sufficient number of images of separate aliquot samples are gathered to provide a

suitable total number of particles with respect to the type of distribution, number or volume based,

and the width of the particle size distribution (see ISO 14488) and that they contain an adequate

statistical number of the largest particle of the target material (see Annex A).
© ISO 2014 – All rights reserved 5
---------------------- Page: 13 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)

f) Some implementations of the image analysis technique employ a large area X, Y servo or manually

controlled sample slide assembly. Such large slides enable many measurement frames of the particles

deposited to be examined. Should the method of fully separate measurement frames be employed

then any frame overlap shall be avoided. If the method of overlapping measurement frames, or

other methods of analysing particles that interact with the measurement frame edge is used, then

procedures shall be employed to ensure that each particle is only included into the total count for

the appropriate size class, once. For more information, see 8.3.

g) If appropriate, ensure that the image quality consisting of illumination, focus and magnification has

not changed at the end of the measurement. The requirement for this step depends on the variability

of the instrument employed.

h) For the case when image analysis is to be used for certified reference material measurement at the

end of the image gathering procedure, the calibration outlined in a) should be repeated and any

measured deviation recorded.

i) All the conditions, set up or established, for the target material shall be fully documented.

5 Sample preparation demands for method description
5.1 Sample splitting and reduction

As only a small amount of material is needed to prepare a test sample, this should be sub-divided from

the whole sample in a manner that ensures that the test sample is representative of the whole as specified

in ISO 14488.
5.2 Touching particles

In order to assess the degree of touching particles, a suitable optical resolution setting of the imaging

system should be chosen. The optical resolution should also meet the criteria set out in 4.2 b).

The number of particles touching each other should be minimized. It is a prime requirement of the

method that measurements shall be made on isolated particles. Touching particles measured as one

particle without a proper separation will introduce error.

It is often not possible to reliably detect touching particles by image analysis alone, but the influence

of touching particles on the result can be investigated experimentally by increasing or decreasing the

number of particles per image. If the number of particles cannot be changed, the influence on the results

can be investigated using a reference material with similar size and shape.
5.3 Particle distribution

There should be an adequate distribution of particles in the field of view. It may be necessary to examine

several fields of view if a large total particle count is required. The whole area of the measurement

field should be examined to ascertain whether there is noticeable segregation of particles (by size). The

requirements set out in 4.2 f) should be followed.
5.4 Number of particles to be counted

The number of particles to be analysed depends upon whether the final result is a particle size

distribution by the number or by the volume of particles. Considerable care has to be exercised in order

to ensure that the analysis is representative of the bulk sample as described in ISO 14488. This can be

demonstrated by splitting the bulk sample into at least three test samples. Each test sample should

contain sufficient particle numbers for a full measurement. Statistical analysis of the data will reveal the

repeatability of the method including sampling and dispersion.
NOTE See Annex A for more information.
6 © ISO 2014 – All rights reserved
---------------------- Page: 14 ----------------------
SIST ISO 13322-1:2015
ISO 13322-1:2014(E)
5.5 Particle suspending fluid

It is likely that a large number of particle measurements will be of particles presented in a gas where an

adequate image contrast should be ensured. Should particle presentation require a liquid suspension

then it is preferred that such liquids be clean, particle-free, transparent and have a refractive index

as different as possible from the refractive index of particles to enhance the image contrast. Particles

presented in a mixed optical background such as in biological specimens, may require dynamic particle

by particle threshold selection.

WARNING — Automated particle by particle parameter selection applied for segmentation as

envisaged when mixed optical backgrounds are required cannot be validated for true particle

size and may result in particle size bias and reduced accuracy.

CAUTION — Particle systems of mixed optical properties may have inaccurate particle sizes

attributed to them due to possible threshold setting errors introducing particle size bias.

6 Quality of captured images
6.1 General

It is important for the analysis that the particles in the captured images are well dispersed. The number

of overlapping or touching particles reduces with the concentration of the particles in the frame (see

5.2). This is in conflict with the requirements of having a large particle
...

Questions, Comments and Discussion

Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.