ISO 19056-3:2022

Microscopes — Definition and measurement of illumination properties — Part 3: Incident light fluorescence microscopy with incoherent light sources

This document specifies procedures for the measurement of illumination brightness, temporal stability and uniformity for incident light fluorescence microscopy. The measurement for uniformity is defined in image planes or intermediate image planes only, when these planes are suitable for detection by electronic imaging devices. This document defines how illumination brightness, temporal stability and uniformity are measured, and how this information is provided to the user. NOTE The scope is intentionally limited to electronic imaging devices and (intermediate) image planes. The visual observation by means of eyepieces would require a different measurement procedure and hence result in ambiguities in the description of measurement procedures. Nevertheless, this document will give useful estimates for the uniformity with visual observation as in this case an eyepiece is used to observe an intermediate image plane (which is under the scope of this document).

Microscopes — Définition et mesurage des propriétés d'éclairage — Partie 3: Microscopie par fluorescence à lumière incidente avec sources lumineuses incohérentes

General Information

• Status: Published
• Publication Date: 09-Jan-2022
• ICS: 37.020 - Optical equipment
• Technical Committee: ISO/TC 172/SC 5 - Microscopes and endoscopes
• Drafting Committee: ISO/TC 172/SC 5/WG 9 - Optical performance of microscope components
• Current Stage: 6060 - International Standard published
• Start Date: 10-Jan-2022
• Due Date: 25-Apr-2022
• Completion Date: 10-Jan-2022

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 19056-3
First edition
2022-01
Microscopes — Definition and
measurement of illumination
properties —
Part 3:
Incident light fluorescence microscopy
with incoherent light sources
Microscopes — Définition et mesurage des propriétés d'éclairage —
Partie 3: Microscopie par fluorescence à lumière incidente avec
sources lumineuses incohérentes
Reference number
ISO 19056-3:2022(E)
© ISO 2022

---------------------- Page: 1 ----------------------
ISO 19056-3:2022(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2022
All rights reserved. Unless otherwise specified, or required in the context of its implementation, 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
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
  © ISO 2022 – All rights reserved

---------------------- Page: 2 ----------------------
ISO 19056-3:2022(E)
Contents  Page
Foreword .iv
Introduction .v
1 Scope . 1
2  Normative references . 1
3  Terms and definitions . 1
4  Measurands . 1
4.1 General . 1
4.2 Illumination brightness . 1
4.3 Temporal stability . 2
4.3.1 General . 2
4.3.2 Short-term stability of radiant flux . 2
4.3.3 Long term stability of radiant flux . 2
4.4 Uniformity . 3
5  Measurement procedure . 3
5.1 General . 3
5.2 Microscope settings . 3
5.3 Illumination brightness . 4
5.4 Temporal stability . 4
5.5 Uniformity . 5
5.6 Spectral information . 6
6  Information provided to the user .6
Annex A (informative) Examples . 7
iii
© ISO 2022 – All rights reserved

---------------------- Page: 3 ----------------------
ISO 19056-3:2022(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 of the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to
the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see
www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 172 Optics  and  photonics,
Subcommittee SC 5, Microscopes and endoscopes.
A list of all parts in the ISO 19056 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.
iv
  © ISO 2022 – All rights reserved

---------------------- Page: 4 ----------------------
ISO 19056-3:2022(E)
Introduction
This document defines important quantities and methods to assess the quality of incident light
fluorescence illumination for microscopes as well as methods to measure them. Details covered will
enable users to compare different microscopes quantitatively with respect to their fluorescence
illumination prior to a purchase decision and chose an appropriate instrument according to their
application. Data obtained by following these methods also enables users to compare existing
instruments with each other or to track performance variations over time, assuring proper performance.
Additionally, what information is to be provided to the user is covered.
v
© ISO 2022 – All rights reserved

---------------------- Page: 5 ----------------------
INTERNATIONAL STANDARD ISO 19056-3:2022(E)
Microscopes — Definition and measurement of
illumination properties —
Part 3:
Incident light fluorescence microscopy with incoherent
light sources
1 Scope
This document specifies procedures for the measurement of illumination brightness, temporal stability
and uniformity for incident light fluorescence microscopy. The measurement for uniformity is defined
in image planes or intermediate image planes only, when these planes are suitable for detection by
electronic imaging devices.
This document defines how illumination brightness, temporal stability and uniformity are measured,
and how this information is provided to the user.
NOTE The scope is intentionally limited to electronic imaging devices and (intermediate) image planes.
The visual observation by means of eyepieces would require a different measurement procedure and hence
result in ambiguities in the description of measurement procedures. Nevertheless, this document will give
useful estimates for the uniformity with visual observation as in this case an eyepiece is used to observe an
intermediate image plane (which is under the scope of this document).
2  Normative references
There are no normative references in this document.
3  Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
4  Measurands
4.1  General
For obtaining the desired image quality, when using a light microscope, the brightness, temporal
stability and uniformity of the image play an important role. This holds true for different applications
and various types of instruments. The measurement of illumination brightness and uniformity is
mandatory. The temporal stability is an optional measurement because it is only suitable for certain
types of light sources (e.g. arc lamps).
4.2  Illumination brightness
The illumination shall be bright enough to allow the detection of the details of the object under
investigation.
1
© ISO 2022 – All rights reserved

---------------------- Page: 6 ----------------------
ISO 19056-3:2022(E)
As this document is based on measurement procedures, the image brightness shall be expressed in
the corresponding SI unit. The radiometric unit irradiance shall be used because it is well suited for
the essentially monochromatic or narrow-band illumination spectral range used in fluorescence
microscopy.
The value is expressed as given by Formula (1):
P
E= (1)
A
where
2
E is the irradiance averaged over the area of the diaphragm used in W/mm ;
P is the radiant flux recorded by the power meter in W;
2
A is the area of diaphragm used in mm .
4.3  Temporal stability
4.3.1  General
The temporal stability of the radiant flux is an important factor when conducting experiments in a time
series. If experimental results are to be derived from variations in the image brightness, any variation
in illumination brightness due to temporal instability shall not contribute to these results.
The temporal stability shall be expressed for at least one of two different time scales.
4.3.2  Short-term stability of radiant flux
The radiant flu
...

FINAL
INTERNATIONAL ISO/FDIS
DRAFT
STANDARD 19056-3
ISO/TC 172/SC 5
Microscopes — Definition and
Secretariat: DIN
measurement of illumination
Voting begins on:
2021-10-19 properties —
Voting terminates on:
Part 3:
2021-12-14
Incident light fluorescence microscopy
with incoherent light sources
Microscopes — Définition et mesurage des propriétés d'éclairage —
Partie 3: Microscopie par fluorescence à lumière incidente avec
sources lumineuses incohérentes
RECIPIENTS OF THIS DRAFT ARE INVITED TO
SUBMIT, WITH THEIR COMMENTS, NOTIFICATION
OF ANY RELEVANT PATENT RIGHTS OF WHICH
THEY ARE AWARE AND TO PROVIDE SUPPOR TING
DOCUMENTATION.
IN ADDITION TO THEIR EVALUATION AS
Reference number
BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO-
ISO/FDIS 19056-3:2021(E)
LOGICAL, COMMERCIAL AND USER PURPOSES,
DRAFT INTERNATIONAL STANDARDS MAY ON
OCCASION HAVE TO BE CONSIDERED IN THE
LIGHT OF THEIR POTENTIAL TO BECOME STAN-
DARDS TO WHICH REFERENCE MAY BE MADE IN
NATIONAL REGULATIONS. © ISO 2021

---------------------- Page: 1 ----------------------
ISO/FDIS 19056-3:2021(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2021
All rights reserved. Unless otherwise specified, or required in the context of its implementation, 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
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
  © ISO 2021 – All rights reserved

---------------------- Page: 2 ----------------------
ISO/FDIS 19056-3:2021(E)
Contents  Page
Foreword .iv
Introduction .v
1 Scope . 1
2  Normative references . 1
3  Terms and definitions . 1
4  Measurands . 1
4.1 General . 1
4.2 Illumination brightness . 1
4.3 Temporal stability . 2
4.3.1 General . 2
4.3.2 Short-term stability of radiant flux . 2
4.3.3 Long term stability of radiant flux . 2
4.4 Uniformity . 3
5  Measurement procedure . 3
5.1 General . 3
5.2 Microscope settings . 3
5.3 Illumination brightness . 4
5.4 Temporal stability . 4
5.5 Uniformity . 5
5.6 Spectral information . 6
6  Information provided to the user .6
Annex A (informative) Examples . 7
iii
© ISO 2021 – All rights reserved

---------------------- Page: 3 ----------------------
ISO/FDIS 19056-3:2021(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 of the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to
the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see
www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 172 Optics  and  photonics,
Subcommittee SC 5, Microscopes and endoscopes.
A list of all parts in the ISO 19056 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.
iv
  © ISO 2021 – All rights reserved

---------------------- Page: 4 ----------------------
ISO/FDIS 19056-3:2021(E)
Introduction
This document defines important quantities and methods to assess the quality of incident light
fluorescence illumination for microscopes as well as methods to measure them. Details covered will
enable users to compare different microscopes quantitatively with respect to their fluorescence
illumination prior to a purchase decision and chose an appropriate instrument according to their
application. Data obtained by following these methods also enables users to compare existing
instruments with each other or to track performance variations over time, assuring proper performance.
Additionally, what information is to be provided to the user is covered.
v
© ISO 2021 – All rights reserved

---------------------- Page: 5 ----------------------
FINAL DRAFT INTERNATIONAL STANDARD ISO/FDIS 19056-3:2021(E)
Microscopes — Definition and measurement of
illumination properties —
Part 3:
Incident light fluorescence microscopy with incoherent
light sources
1 Scope
This document specifies procedures for the measurement of illumination brightness, temporal stability
and uniformity for incident light fluorescence microscopy. The measurement for uniformity is defined
in image planes or intermediate image planes only, when these planes are suitable for detection by
electronic imaging devices.
This document defines how illumination brightness, temporal stability and uniformity are measured,
and how this information is provided to the user.
NOTE The scope is intentionally limited to electronic imaging devices and (intermediate) image planes.
The visual observation by means of eyepieces would require a different measurement procedure and hence
result in ambiguities in the description of measurement procedures. Nevertheless, this document will give
useful estimates for the uniformity with visual observation as in this case an eyepiece is used to observe an
intermediate image plane (which is under the scope of this document).
2  Normative references
There are no normative references in this document.
3  Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
4  Measurands
4.1  General
For obtaining the desired image quality, when using a light microscope, the brightness, temporal
stability and uniformity of the image play an important role. This holds true for different applications
and various types of instruments. The measurement of illumination brightness and uniformity is
mandatory. The temporal stability is an optional measurement because it is only suitable for certain
types of light sources (e.g. arc lamps).
4.2  Illumination brightness
The illumination shall be bright enough to allow the detection of the details of the object under
investigation.
1
© ISO 2021 – All rights reserved

---------------------- Page: 6 ----------------------
ISO/FDIS 19056-3:2021(E)
As this document is based on measurement procedures, the image brightness shall be expressed in
the corresponding SI unit. The radiometric unit irradiance shall be used because it is well suited for
the essentially monochromatic or narrow-band illumination spectral range used in fluorescence
microscopy.
The value is expressed as given by Formula (1):
P
E= (1)
A
where
2
E is the irradiance averaged over the area of the diaphragm used in W/mm ;
P is the radiant flux recorded by the power meter in W;
2
A is the area of diaphragm used in mm .
4.3  Temporal stability
4.3.1  General
The temporal stability of the radiant flux is an important factor when conducting experiments in a time
series. If experimental results are to be derived from variations in
...