SIST-TS CEN/TS 16827-1:2015
(Main)Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for FFPE tissue - Part 1: Isolated RNA
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for FFPE tissue - Part 1: Isolated RNA
This Technical Specification recommends the handling, documentation and processing of FFPE tissue specimens intended for RNA analysis during the preanalytical phase before a molecular assay is performed. This Technical Specification is applicable to molecular in vitro diagnostic examinations (e.g., in vitro diagnostic laboratories, laboratory customers, in vitro diagnostics developers and manufacturers, institutions and commercial organizations performing biomedical research, biobanks, and regulatory authorities).
The formalin-fixation and the paraffin embedding process lead to modifications of the RNA molecules, which can impact the validity and reliability of the analytical test results.
Therefore, special measures have to be taken to minimize the described profile changes and modifications within the tissue for subsequent RNA analysis.
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für präanalytische Prozesse für FFPE-Gewebe - Teil 1: Isolierte RNS
Diese Technische Spezifikation gibt Empfehlungen zur Handhabung, Dokumentation und Verarbeitung von aus FFPE Gewebe bestehendem und für die RNS Analyse vorgesehenem Untersuchungsmaterial während der präanalytischen Phase vor Beginn der molekularen Analyse. Diese Technische Spezifikation gilt für molekulare in vitro diagnostische Untersuchungen (z. B. In vitro Diagnostik Labore, Kunden dieser Labore, Entwickler und Hersteller von In vitro Diagnostika, Einrichtungen und kommerzielle Organisationen, die in der biomedizinischen Forschung tätig sind, Biobanken und Aufsichtsbehörden).
Die Fixierung mit Formalin und die Einbettung in Paraffin können zu Modifikationen der RNS Moleküle führen, die sich auf die Gültigkeit und Zuverlässigkeit der Ergebnisse analytischer Prüfungen auswirken können.
Daher ist es wichtig, dass besondere Maßnahmen getroffen werden, um die beschriebenen Profil-veränderungen und modifikationen im Gewebe im Hinblick auf die anschließende RNS Analyse möglichst gering zu halten.
Tests de diagnostic moléculaire in vitro - Spécifications relatives aux processus préanalytiques pour les tissus FFPE - Partie 1: ARN extrait
La présente Spécification technique fournit des recommandations pour la manipulation, la documentation et le traitement d’échantillons primaires de tissus FFPE destinés à l’analyse de l’ARN durant la phase préanalytique précédant la réalisation d’un essai moléculaire. La présente Spécification technique s’applique aux examens de diagnostic moléculaire in vitro (par exemple, laboratoires de diagnostic in vitro, clients de laboratoires, concepteurs et fabricants de diagnostic in vitro, institutions et organisations commerciales de recherche biomédicale, biobanques et autorités réglementaires).
Le processus de fixation au formol et d'inclusion en paraffine conduit à des modifications des molécules d'ARN, ce qui peut avoir un impact sur la validité et la fiabilité des résultats d'essais analytiques.
Il est donc essentiel de prendre des mesures spéciales afin de réduire le plus possible les changements de profil décrits et les modifications dans le tissu lors de l'analyse subséquente de l’ARN.
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za FFPE tkiva - 1. del: Izolirani RNK
Ta tehnična specifikacija vsebuje priporočila za obravnavo, dokumentiranje in obdelavo vzorcev tkiv FFPE, namenjenih za analizo RNK med predanalizno fazo, preden se izvede molekularni preskus. Ta tehnična specifikacija se uporablja za molekularne diagnostične preiskave in vitro (npr. diagnostični laboratoriji in vitro, laboratorijske stranke, razvijalci in proizvajalci diagnostike in vitro, institucije in komercialne organizacije, ki izvajajo biomedicinske raziskave, biobanke ter regulativni organi).
Fiksacija v formalinu in postopek vklopa v parafin vodita do sprememb molekul RNK, kar lahko vpliva na veljavnost in zanesljivost rezultatov analitičnih preskusov.
Zato je treba sprejeti posebne ukrepe, da se zmanjšajo opisane spremembe profila v tkivu za nadaljnje analize RNK.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TS CEN/TS 16827-1:2015
01-oktober-2015
0ROHNXODUQHGLDJQRVWLþQHSUHLVNDYHLQYLWUR6SHFLILNDFLMH]DSUHGSUHLVNRYDOQH
SURFHVH]D))3(WNLYDGHO ,]ROLUDQL51.
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes
for FFPE tissue - Part 1: Isolated RNA
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für FFPE-Gewebe - Teil 1: Isolierte RNS
Tests de diagnostic moléculaire in vitro - Spécifications relatives aux processus
préanalytiques pour les tissus FFPE - Partie 1: ARN extrait
Ta slovenski standard je istoveten z: CEN/TS 16827-1:2015
ICS:
11.100.10 'LDJQRVWLþQLSUHVNXVQL In vitro diagnostic test
VLVWHPLLQYLWUR systems
SIST-TS CEN/TS 16827-1:2015 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST-TS CEN/TS 16827-1:2015
TECHNICAL SPECIFICATION
CEN/TS 16827-1
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
August 2015
ICS 11.100.10
English Version
Molecular in vitro diagnostic examinations - Specifications for
pre-examination processes for FFPE tissue - Part 1: Isolated
RNA
Tests de diagnostic moléculaire in vitro - Spécifications Molekularanalytische in-vitro-diagnostische Verfahren -
relatives aux processus préanalytiques pour les tissus Spezifikationen für präanalytische Prozesse für FFPE-
FFPE - Partie 1: ARN extrait Gewebeproben - Teil 1: Isolierte RNS
This Technical Specification (CEN/TS) was approved by CEN on 6 July 2015 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2015 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16827-1:2015 E
worldwide for CEN national Members.
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Contents Page
European foreword .3
Introduction .4
1 Scope .5
2 Normative references .5
3 Terms and definitions .5
4 General considerations .7
5 Outside the laboratory .8
5.1 Primary tissue collection manual.8
5.1.1 Information about the primary sample donor .8
5.1.2 Information on the primary tissue sample .8
5.1.3 Information on the primary tissue sample processing .8
5.2 Transport requirements .9
6 Inside the laboratory .9
6.1 Information on the primary tissue sample receipt .9
6.2 Formalin fixation of the specimen .9
6.3 Evaluation of the pathology of the specimen and selection of the sample. 10
6.4 Post-fixation of frozen samples . 11
6.5 Processing and paraffin embedding. 11
6.6 Storage requirements . 12
6.7 Isolation of the total RNA . 12
6.7.1 General . 12
6.7.2 General information for RNA isolation procedures . 12
6.7.3 Using commercial kits . 13
6.7.4 Using the laboratories’ own protocols . 13
6.8 Quantity and quality assessment of isolated RNA . 14
6.9 Storage of isolated RNA . 14
Annex A (informative) Quality control of RNA extracted from formalin fixed and paraffin
embedded tissue samples: implications for RT-qPCR based analyses . 15
A.1 Summary . 15
A.2 Results . 15
A.2.1 Time dependency of RNA integrity . 15
A.2.2 Impact of formalin-fixation on cDNA synthesis efficiency . 16
A.2.3 Fixation and storage introduces major gene-to-gene variations in RT-qPCR . 17
A.2.4 Impact of storage conditions of FFPE blocks on RNA Integrity . 18
A.3 Conclusions . 18
A.4 Further reading . 19
Bibliography . 20
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European foreword
This document (CEN/TS 16827-1:2015) has been prepared by Technical Committee CEN/TC 140 “In vitro
diagnostic medical devices”, the secretariat of which is held by DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany,
Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
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Introduction
Molecular in vitro diagnostics has enabled a significant progress in medicine. Further progress is expected by
new technologies analysing signatures of nucleic acids, proteins, and metabolites in human tissues and body
fluids. However, the profiles and/or integrity of these molecules can change drastically during primary sample
collection, transport, storage, and processing thus making the outcome from diagnostics or research
unreliable or even impossible because the subsequent analytical assay will not determine the situation in the
patient but an artificial profile generated during the pre-examination process. Therefore, a standardization of
the entire process from primary sample collection to RNA analysis is needed. Studies have been undertaken
to determine the important influencing factors. This Technical Specification draws upon such work to codify
and standardize the steps for formalin fixed and paraffin embedded (FFPE) tissue with regard to RNA analysis
in what is referred to as the preanalytical phase.
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1 Scope
This Technical Specification gives recommendations for the handling, documentation and processing of FFPE
tissue specimens intended for RNA analysis during the preanalytical phase before a molecular assay is
performed. This Technical Specification is applicable to molecular in vitro diagnostic examinations (e.g.,
in vitro diagnostic laboratories, laboratory customers, developers and manufacturers of in vitro diagnostics,
institutions and commercial organizations performing biomedical research, biobanks, and regulatory
authorities).
The formalin fixation and the paraffin embedding process lead to modifications of the RNA molecules, which
can impact the validity and reliability of the analytical test results.
Therefore, it is essential to take special measures to minimize the described profile changes and modifications
within the tissue for subsequent RNA analysis.
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.
EN ISO 15189:2012, Medical laboratories — Requirements for quality and competence (ISO 15189:2012,
Corrected version 2014-08-15)
ISO 15190, Medical laboratories — Requirements for safety
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN ISO 15189:2012 and the following
apply.
3.1
ambient temperature
unregulated temperature of the surrounding air
3.2
analytical phase
processes that start with the isolated analyte and include all kinds of parameter testing or chemical
manipulation for quantitative or qualitative analysis
3.3
cold ischemia
condition after removal of the tissue from the body until its stabilization or fixation
3.4
FFPE
formalin fixation and paraffin embedding
3.5
FFPE tissues
formalin fixed and paraffin embedded tissues
3.6
formalin
saturated formaldehyde solution containing a mas fraction of 37 % (corresponding to a volume fraction of 40
%) formaldehyde, termed 100 % formalin
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3.7
formalin fixation
treatment of a sample with standard buffered formalin solution for stabilization
3.8
pre-examination processes
preanalytical phase
preanalytical workflow
processes that start, in chronological order, from the clinician’s request and include the examination request,
preparation and identification of the patient, surgical procedure, collection of the primary sample(s), temporary
storage, transportation to and within the analytical laboratory, aliquoting, retrieval, isolation of analytes, and
end when the analytical examination begins
[SOURCE: EN ISO 15189:2012, definition 3.15, modified — An additional term was added and more details
were included.]
Note 1 to entry: The preanalytical phase may include preparative processes that may influence the outcome of the
intended examination.
3.9
primary sample
specimen
discrete portion of a body fluid, breath, hair or tissue taken for examination, study or analysis of one or more
quantities or properties assumed to apply for the whole
[SOURCE: EN ISO 15189:2012, 3.16, modified — The term and definition is used here without the original
notes.]
3.10
quantitative RNA profile
RNA profile
amounts of the individual RNA molecules that are present in a sample and that can be measured in the
absence of any losses, inhibition and interference
3.11
RNA
ribonucleic acid
polymer of ribonucleotides occurring in a double-stranded or single-stranded form
[SOURCE: EN ISO 22174:2005, 3.1.3]
3.12
room temperature
temperature which is defined as 18 °C to 25 °C for the purposes of this document
3.13
sample
one or more parts taken from a primary sample
[SOURCE: EN ISO 15189:2012, 3.24, modified — The example was not taken over.]
3.14
stability
ability of a sample material, when stored under specified conditions, to maintain a stated property value within
specified limits for a specified period of time
[SOURCE: ISO Guide 30:1992, 2.7]
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Note 1 to entry: The measured constituent for the purpose of this document is RNA.
3.15
standard buffered formalin solution
10 % formalin solution containing a mass fraction of 3,7 % (corresponding to a volume fraction of 4 %)
formaldehyde buffered to pH 6,8 to pH 7,2
Note 1 to entry: Standard buffered formalin solutions often contain methanol to inhibit oxidation and polymerization of
formaldehyde.
3.16
warm ischemia
warm Ischemia is the condition where the tissue is deprived of its normal blood supply containing oxygen and
nutrients while the tissue is at body temperature
4 General considerations
For general statements on primary sample collection and handling (including avoidance of cross
contaminations) see EN ISO 15189:2012, 5.4.4, 5.2.6. Consumables including kits shall be verified before use
in examination (see EN ISO 15189:2012, 5.3.2.3); EN ISO 15189:2012, 5.5.1.2 and 5.5.1.3 can also apply.
As all steps of a diagnostic workflow can influence the final analytical performance, the entire workflow
comprising the preanalytical steps, including information on biomolecule stability and storage conditions, and
analytical steps should be verified and validated (see EN ISO 15189).
The stability of the specific quantitative RNA profile(s) of interest should be investigated throughout the entire
preanalytical workflow prior to the development and implementation of an analytical test.
Before tissues are fixed in standard buffered formalin solution, RNA profiles can change significantly
depending on the duration of warm and cold ischemia and the temperature before formalin fixation (e.g., gene
induction, gene down regulation, RNA degradation). In addition, those changes can vary in tissues from
different donors / patients.
Generally, the longer the warm and cold ischemia times and the higher the ambient temperature before
fixation of the tissue specimen, the higher is the risk that changes in the RNA profile can occur.
NOTE Intraoperative warm ischemia can result in more pronounced changes of RNA profiles than in postoperative
cold ischemia. RNA profiles can also vary, depending on the origin and type of tissue, the underlying disease, the surgical
procedure, drugs administered for anaesthesia or treatment of concomitant disease, and on the different environmental
conditions after the tissue removal from the body.
As warm ischemia cannot be easily standardized, its time and duration should be documented. When it is not
possible to avoid cold ischemia, its time of onset and duration shall be documented and the temperatures of
the specimen transport container’s surroundings should be documented. Where the specimen is transported
to another facility for formalin fixation, the transport duration shall be documented and the ambient conditions
should also be documented.
In addition, formalin fixation causes modifications of biomolecules and leads to suboptimal performance of
RNA extracted from FFPE tissues that should be considered in quality control and application of molecular
assays, especially in the context of gene expression studies (see [1], [2], [3]). Assay optimization for FFPE
tissues or the use of non-crosslinking alternatives to standard buffered formalin solution are options to
minimize this issue for molecular analyses (see [4]).
Safety regulations on transport and handling shall be considered (see EN ISO 15189:2012, 5.2.3 and 5.4.5
and ISO 15190).
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During the whole preanalytical workflow precautions shall be taken to avoid cross contamination between
different samples.
If a commercial product is not used in accordance with the manufacturers' instructions, responsibility for its
use and performance lies with the user.
5 Outside the laboratory
5.1 Primary tissue collection manual
5.1.1 Information about the primary sample donor
The documentation should include, but is not limited to:
a) the primary donor / patient ID, which can be in the form of a code;
b) the health status of the primary sample donor (e.g., healthy, disease type, concomitant disease);
c) the information about routine medical treatment and special treatment prior to tissue collection (e.g.,
anaesthetics, medications, surgical or diagnostic procedures (e.g., biopsy device used for the collection));
d) the start of ischemia within the body (warm ischemia) by documenting the ischemia-relevant vessel
ligation/clamping time point (usually arterial clamping time).
5.1.2 Information on the primary tissue sample
The documentation shall include, but is not limited to:
a) the time point when tissue is removed from the body;
b) the description of tissue type, tissue condition (e.g., diseased, unaffected by the disease) and organ
tissue of origin, including references to any marking applied in the operating theatre made by surgeon,
radiologist or pathologist;
c) the documentation steps described under 6.2, if the formalin fixation starts outside the laboratory.
5.1.3 Information on the primary tissue sample processing
The following steps shall be performed:
1. the documentation of any additions or modifications to the primary sample after removal from the body
(e.g., labelling for the orientation of the specimen (e.g., ink-marking, stitches), incision(s));
2. the selection and use of transport containers and packages (e.g., cooling box, box for storing and
transportation, vacuum packaging) fit for transport of formalin fixed tissue samples, if relevant;
3. the selection and use of stabilization procedures (e.g., cooling methods) for transport;
NOTE 1 Accidentally freezing and thawing the tissue (e.g., by using cool packs in a wrong manner) can lead to
RNA degradation when the tissue thaws thereafter. It can also impact the morphological characterization.
NOTE 2 This step can be omitted, if the specimen is transferred directly into standard buffered formalin solution
(see 6.2).
4. the labelling of the transport container (e.g., registration-number, barcode (1D or 2D), primary sample
type, quantity, and organ tissue of origin) and additional documentation (information as specified in 5.1.1,
5.1.2, and 5.1.3, 1. to 3.). If a single sample container contains several aliquots of the same specimen,
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and the aliquots represent different features (e.g., tissue type, disease status, location) this shall be
documented.
Specimens should be transferred without delay into the transport container after the removal from the body.
The container should then be kept on wet-ice or at 2 °C to 8 °C in order to minimize RNA profile changes.
The temperatures of the transport container's surroundings during cold ischemia time (e.g., temperatures in
different rooms, transport) should be documented. If the temperature cannot be measured, the temperature
range should be estimated by classification as ambient temperature, room temperature, or at 2 °C to 8 °C.
5.2 Transport requirements
The laboratory in partnership with the clinical or surgery department shall establish a protocol for the transport
procedure of the specimen.
If the primary tissue sample is not already placed into standard buffered formalin solution, it should be
transported on wet-ice or at 2 °C to 8 °C without delay in order to minimize the changes to the RNA profile.
NOTE There is evidence that RNA in tissues can be stabilized in plastic bags under vacuum when kept at 0 °C to
4 °C during transport [5] before the samples are archived for biobanks or used for histopathological evaluation.
If the primary tissue sample is already placed into standard buffered formalin solution outside the laboratory,
the temperature during transport should not exceed room temperature.
The compliance with the protocol for the transport procedure shall be documented. Any deviations from the
protocol shall be described and documented.
6 Inside the laboratory
6.1 Information on the primary tissue sample receipt
The name of the person receiving the primary tissue sample shall be documented. The tissue sample arrival
time and conditions (e.g., labelling, transport conditions including temperature, tissue type and quantity of the
primary sample, leaking/breaking of the container) of the received samples shall be documented. Any
deviations from the established protocol for the transport procedure (see 5.2) shall be documented.
NOTE Temperature conditions during transport can influence the quantitative RNA profile and RNA quality.
6.2 Formalin fixation of the specimen
The fixative used shall be standard buffered formalin solution.
The pH-value and concentration of the standard buffered formalin solution should be checked regularly as
formalin is not stable (e.g., it has a tendency to be oxidized to formic acid [6]).
The following steps shall be performed:
1. the consultation with the manufacturer's material safety data sheet (MSDS) before handling standard
buffered formalin solution;
NOTE Formaldehyde is a hazardous compound that chemically modifies biomolecules.
2. the documentation of the time point of placing the tissue sample into standard buffered formalin solution;
NOTE The total formalin fixation duration can impact further analyses e.g., immunohistochemical techniques,
nucleic acid based molecular analyses [3]; see also A.2.1 and A.2.2. The optimal formalin fixation duration can vary
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depending on tissue type and size. For larger surgical specimen inhomogeneous fixation can occur before the
grossing process due to slow penetration of formaldehyde from the surface of the tissue to the interior.
EXAMPLE For tissue pieces with a maximum thickness of 5 mm fixation times between 12 h and 24 h are
reasonable in most cases. See also 6.7.2.
3. the selection of collection container/s:
a) the capacity of the collection containers should be such that the primary sample can be completely
submerged into the standard buffered formalin solution. The minimum standard buffered formalin
solution to tissue ratio depends on the tissue concerned, but should be at least 4:1, (volume to
volume). To ensure complete formalin fixation of larger specimens a special tissue handling such as
incision(s) of solid organs or opening of hollow organs can be necessary. -
NOTE The fixative-to-tissue-volume-ratio influences the fixation progression. There is evidence that a ratio of 10:1
can be necessary [7]. If the container has a small volume, standard buffered formalin solution can be changed
periodically, because formalin reacts with tissue components in a time-dependent way.
b) when using containers pre-filled with standard buffered formalin solution, provider’s product
instructions shall be followed;
c) the container shall be securely closable;
4. the labelling of the container (e.g., by using self-adhesive labels, handwriting, radio frequency
identification devices (RFID), pre-labelled containers, bar codes) shall give the minimum information of:
a) the patient ID, which can be in the form of a code;
b) the basic information on e.g., the tissue type, tissue condition, and related additional information such
as affected (e.g., tumour) or unaffected, unless a sample tracking system can supply this information
coupled to the identification of the sample used in 6.2, 4. a);
c) the unique numbering of each container;
5. the documentation of types, quantity and description of samples.
It should be considered that under some disease conditions, such as tumours, molecular features may not be
present homogeneously in the tissue sample. Therefore, it is important that the part of the actual tissue
sample used for molecular analysis is evaluated by a medically qualified (e.g., board certified) pathologist. In
this context it should be documented which features of a disease are actually reflected in the tissue sample
used for molecular analysis (e.g., different molecular mechanisms can be activated at the centre and at the
invasion front of the tumour; also tumours can be composed of areas showing different differentiation grades).
6.3 Evaluation of the pathology of the specimen and selection of the sample
The evaluation and documentation of the pathology of the specimen and the selection of the sample from the
specimen for further processing shall be done by or under supervision or responsibility of a medically qualified
(e.g., board certified) pathologist.
Local, national or regional regulations may apply.
Options to select the sample for RNA analysis:
a) The selection of appropriate parts of the specimen for molecular analyses and histopathological analyses
as well as for optional further research purposes shall be done by or under supervision of a medically
qualified (e.g., board certified) pathologist to ensure that the collection of the sample for RNA analysis
does not compromise the histopathological analyses.
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In the context of macroscopic evaluation of the surgical specimen before and/or after formalin fixation, the
clinical instructions, number, name of the patient, date of birth of the patient and type of tissue shall be
checked. The surgical specimen and all findings shall be described appropriately according to the
guidelines of the respective medical societies and in correlation with the clinical instructions and
questions. The anatomic localization represented in the specimen shall be described, resection margins
and other important areas may be marked if necessary and be helpful for later microscopic evaluation;
photographs may be taken. Where representative samples for microscopic evaluation are required (i.e.,
grossing) this shall be done in accordance with the organ/disease-specific guidelines from the respective
medical societies.
b) Where the tissue specimen was removed from the body without the requirement of histopathological
diagnosis; documentation of this specimen, the evaluation, as well as the documentation and selection of
the samples may be done by other qualified persons than pathologists.
NOTE Documentation can include photographs.
The size of the samples should be appropriate for the tissue cassette (maximum of approximately
3 cm × 2 cm × 0,5 cm). If the specimen is not yet fixed appropriately, post-fixation can be performed within the
tissue cassette. Each tissue cassette shall be labelled
...
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