Bms Training in Immunocytochemistry Logbook for General Certificate of Competence

CERTIFICATE OF EXPERT PRACTICE IN

FLOW CYTOMETRY

Indicative Study Guide

12 Coldbath Square London EC1R 5HL Tel: 020 7713 0214 Fax: 020 7436 4946 Email: [email protected] Website: www.ibms.org TABLE OF CONTENTS

GUIDANCE TO CANDIDATES 3

TRAINING MODULES 8

GENERIC MODULES 8

A1. HEALTH AND SAFETY 8

A2. RISK MANAGEMENT/CLINICAL RISK/CLINICAL GOVERNANCE 8

A3. QUALITY AND AUDIT 9

A4. EQUIPMENT 10

A5. DATA ACQUISITION AND ANALYSIS 10

A6. FILING AND ARCHIVING 10

SPECIALIST MODULES 12

B1. HAEMATOLOGY 12

B2. IMMUNOLOGY 15

B3. HISTOCOMPATABILITY AND IMMUNOGENETICS 17

B4. STEM CELL BIOLOGY 20

B5. TRANSFUSION 22

SUGGESTED READING LIST 24 GUIDANCE TO CANDIDATES

INTRODUCTION The Institute guidance provides a nationally recognised framework to enable biomedical scientists to demonstrate the essential level of competence necessary to perform techniques in flow cytometry and to demonstrate an up-to-date knowledge of diagnostic applications. The training syllabus covers the fundamental preparations and analyses commonly used in laboratories. In addition, an awareness of more specialised areas must be included to form a baseline of practice knowledge.

Laboratories wishing to offer this training must have Institute approval for registration training and be CPA registered. Training is conducted in-house under the overall responsibility of a named appropriate experienced, biomedical scientist or a clinical scientist

Institute’s Examination Structure

HCS Career Membership Framework Stage Class

9 Professional Doctorate

8 Advanced Specialist Diploma

Diplomas of Expert Practice Fellow 7 Higher Specialist Diploma

Certificates of 6 MSc Expert Practice Member Specialist Diploma

5 Licentiate Certificate of Competence

BSc (Hons) Biomedical Science

ELIGIBILITY The undertaking of flow cytometry is a specialised area and requires the acquisition of specific knowledge and skills. The minimum requirements for entry to training for the Certificate of Expert Practice in Flow Cytometry are:  Member of the Institute of Biomedical Science (MIBMS)  Institute of Biomedical Science Specialist Diploma or equivalent  Registered practitioner with the HPC  A minimum of two years whole-time equivalent post-registration laboratory experience which includes flow cytometry”

CEP Flow Cytometry guidance to candidates Nov 10 3  Employment in, or secondment to, a CPA registered laboratory or equivalent with Institute registration training approval  A named supervisor (expert biomedical scientist or clinical scientist)  Approval from the laboratory manager  Training delivered in accordance with the Institute’s guidance.

AIMS 1. To extend the knowledge and skills a candidate has obtained from at least two years post registration experience in an appropriate laboratory, the completion of a relevant specialist diploma or equivalent.

2. To enable successful candidates to demonstrate that they have the required knowledge, skills and competencies that contribute to the investigations relating to flow cytometry at an intermediate level.

LEARNING OUTCOMES Individuals awarded the Certificate of Expert Practice in Flow Cytometry will be able to:

1. Explain scientific, technical and diagnostic issues related to flow cytometry as evidenced in the written examination and the training portfolio.

2. Demonstrate ability to practice in a routine/specialised laboratory with scientific and technical skills to facilitate effective and accurate investigation as evidenced in the training portfolio.

3. Demonstrate a wide understanding of analytical principles and procedural limitations as evidenced in the training portfolio.

4. Demonstrate knowledge and awareness of current guidelines related to the laboratory investigation as evidenced in the training portfolio and the written examination.

MANDATORY TRAINING COURSES It is mandatory that candidates undertake one of the following courses as part of their training programme:

i) Queen Mary University of London two-day flow cytometry course. Course tutor: Professor Marion Macey ii) Royal Microscopical Society – module 1 of the five-day course. iii) Royal Microscopical Society five-day course for the IBMS Certificate of Expert Practice. Course tutor: Dr M Ormerod.

Candidates who have completed the above training courses are eligible to undertake the qualification within to a maximum of five years.

TRAINING LABORATORY Any laboratory or laboratory network wishing to offer support for a biomedical scientist preparing for the Certificate of Expert Practice in Flow Cytometry must have a significant annual through-put of cases. This may represent predominantly one type of tissue or a range of different types of samples.

CEP Flow Cytometry guidance to candidates Nov 10 4 NAMED SUPERVISOR The decision to support any eligible biomedical scientist in this training lies with the individual laboratory manager. The supervisor must have an active interest in flow cytometry. In order to fulfil the training requirements it is acceptable for an arrangement to exist with another hospital for a period of secondment in order to obtain the required level of practical experience and competence in the use of techniques not used in the employing laboratory. All supervisors including those who have taken responsibility for specific aspects of the candidate’s training must complete the ‘Supervisor/mentor details’ section on the candidate’s application form and sign the declaration.

LEARNING STRATEGY Candidates are advised to undertake the following activities to prepare for assessment: 1. Continuing Professional Development (CPD) 2. Reading and reflecting on scientific and technical papers 3. Self-structured reading 4. Attendance at seminars/lectures 5. Undertake training programmes related to investigation.

ASSESSMENT STRUCTURE

PART A – Training portfolio Portfolios should be submitted in hard copy format by the published deadline. The exact date will be published in The Biomedical Scientist and on the Institute’s website. Candidates will not be required to attend the portfolio assessment. The Institute will endeavour to carry out all portfolio assessments within eight weeks of the submission deadline.

Candidates must achieve a pass grade in the portfolio assessment to be able to proceed to Part B. Unsuccessful candidates will be advised on areas which require development, and on the time scale for resubmission.

It is expected that the assessment of competence will be an on-going process throughout the preparation period. The training portfolio should allow for the recording of comments regarding progress and aptitude.

i) Theoretical knowledge and practical skills Each aspect of preparation comprises the theoretical knowledge required to understand the processes that underpin the task and the practical skills and competencies to successfully execute the task. The biomedical scientist will be expected to acquire and demonstrate the knowledge that accompanies the practical skills.

ii) Standard operating procedures All aspects of laboratory work must be covered by individually signed, indexed and dated standard operating procedures (SOPs). Before commencing training it is mandatory that appropriate SOPs be in place to describe the departmental protocols for pre-analytical and analytical phases of flow cytometry including approved minimum data sets. The biomedical scientist must operate within the SOP at all appropriate times.

iii) Quality control and audit Audit forms an integral part of the training process. The requirement for review of cases forms the basis of continuing audit of biomedical scientist’s competence and performance. Documentary evidence of this practice must be kept as part of the training record.

CEP Flow Cytometry guidance to candidates Nov 10 5 The training portfolio must clearly demonstrate the organising and recording of the candidate’s achievements and reflect the range of required competencies, skills and experience. As a minimum, the portfolio must include the following elements along with any additional and appropriate documentation relating to the candidate’s training:  A log of the case repertoire encountered during the training period  Copies of method sheets and standard operating procedures  Evidence of cases reviewed with the supervisor  Copies of attendance certificates at training events and evidence of involvement in training or teaching non-laboratory staff in analytical/diagnostic practice outside the laboratory.

Candidates must complete all generic and common modules along with a choice of two specialist modules as described in the IBMS Certificate of Expert Practice in Flow Cytometry indicative study guide and training portfolio. Candidates may use the training module sheets with the appropriate signed supervisor declarations from this guide as part of their portfolio.

PART B – Written examination (120 minutes) The examination paper will comprise a generic and a discipline specific section. Candidates will be expected to answer two questions from a choice of four in each section.

In the written examination candidates must achieve a combined overall average of 50, with neither of the two papers scoring below 40%.

On securing a pass in both parts of the assessment candidates will be awarded the Institute’s Certificate of Expert Practice in Flow Cytometry.

APPLICATION FORMS Application forms are available from the Institute’s Office using the contact details below and may be requested by telephone or e-mail, or they may be downloaded from the Institute’s web site.

The completed application together with the correct fee must be returned to the Institute.

Incomplete, illegible or applications without fees will be returned for correction and resubmission before acceptance.

CONFIRMATION OF APPLICATION Once accepted, candidates will be sent a confirmation of candidacy and a reminder of the submission deadline for examination portfolios.

DEFERRALS AND WITHDRAWALS Candidates who wish to defer entry to an examination must contact the Institute a minimum of six weeks prior to the date of the examination will be entitled to a full transfer of their fees. Any deferrals made after this deadline will only be entitled to a 50% fee transfer unless proven mitigating circumstances exist. A maximum of two deferrals is permitted.

Candidates wishing to withdraw from an examination at any time will not be entitled to any reimbursement of the examination fee unless proven mitigating circumstances exist.

Candidates who are required to submit a portfolio for reassessment following a referral would be required to pay a reassessment fee.

APPEALS Any candidate wishing to appeal against the outcome of the assessment procedure must contact the examinations department and request an appeals form. This must be completed and returned to the Chief Executive within a maximum period of 40 days, following publication of the results.

CEP Flow Cytometry guidance to candidates Nov 10 6 TRAINING MODULES

SECTION A

GENERIC MODULES

Candidates are expected to complete all modules in this section.

A1. HEALTH AND SAFETY

A1.1. GENERAL PRINCIPLES Understands: A1.1.1. the safety responsibilities of the employee under the Health and Safety at Work Act 1974, COSHH and current safety legislation

A1.1.2. other laws and regulations relating to health, safety and security

A1.1.3. health, safety and security risks and the actions required minimising them

A1.1.4. the departmental safety policy

A1.1.5. the correct use of personal protective equipment (PPE)

A1.1.6. correct use of display screen and equipment.

A1.2. HAZARDS ASSOCIATED WITH CELL / TISSUE PREPARATION Understands: A1.2.1. safe handling of cells or tissue for investigation

A1.2.2. operating of equipment used in sample preparation

A1.2.3. analytical procedures for preparing samples

A1.2.4. the handling of infectious materials

A1.2.5. the local disinfectant, sterilisation and disposal procedures

A1.2.6. the control, storage and disposal of reagents and chemicals

A2. RISK MANAGEMENT/CLINICAL RISK/CLINICAL GOVERNANCE

The requirements for full standard operating procedure (SOP) risk assessment compliance, together with knowledge of risk assessment relating to:

A2.1. POLICIES

A2.1.1. Local procedures for entering specimen into database systems.

A2.1.2. Local procedures for accurate numbering and labelling of specimens.

A2.1.3. Local policy for ‘specimen acceptance’, i.e. minimum data requirements.

A2.1.4. The clinical risk to patients with regard to the mislabelling of samples.

CEP Flow Cytometry guidance to candidates Nov 10 7 A2.1.5. The clinical risk to the patient with regard to the mislabelling of reagents.

A2.1.6. Quality control, quality assurance and quality assessment.

A2.1.7. Patient confidentiality and consent.

A2.1.8. Clinical effectiveness in the timely production of test results.

A2.1.9. Participation in clinical audit.

A2.1.10. On-going staff training and education.

A2.1.11. The contribution of flow cytometry analysis in clinical management.

A2.2. SPECIMEN PROCESSING

A2.2.1. PRE-ANALYSIS

A2.2.1.1. Appropriate specimen and quality for the investigations requested

A2.2.2. ANALYSIS

A2.2.2.1. Is competent in the preparation and correct identification of cells with labelled antibodies appropriate for the clinical question

A2.2.3. POST-ANALYSIS

A2.2.3.1. Is competent in data analysis to accurately identify cell populations present and quantify as appropriate

A3. QUALITY AND AUDIT

A3.1. QUALITY MANAGEMENT Understands the requirement for quality management and has knowledge of: A3.1.1. departmental quality policy

A3.1.2. departmental quality management system

A3.1.3. quality assurance: records and documentation

A3.1.4. quality control

A3.1.5. quality assessments – internal (IQA) and external (EQA) systems

A3.1.6. audit trails

A3.1.7. accreditation by an authoritative body.

A3.2. CLINICAL AUDIT Has awareness of: A3.2.1. the need for audit

A3.2.2. present evidence of participation in audit processes

A3.2.3. types of audit

A3.2.4. audit documents. CEP Flow Cytometry guidance to candidates Nov 10 8 A4. EQUIPMENT

A4.1. FLOW CYTOMETRY Understands the principles of and is competent in the operation and routine maintenance of flow cytometer and use of application software for analyser operation and data analysis to include: A4.1.1. principles and practice of flow cytometry

A4.1.2. set-up for optimal detection conditions according to cell type

A4.1.3. multi-colour analysis

A4.1.4. calibration systems

A4.1.5. trouble-shooting

A5. DATA ACQUISITION AND ANALYSIS

Is competent at:

A5.1. performing measurements applying measurement markers to flow cytometry scatter plots and histograms

A5.2. producing appropriate list mode data files

A5.3. saving data in a variety of formats as required by local procedures

A5.4. data manipulation within a software package.

A5.5. storage of raw data

Understands:

A5.6. the importance of correct patient ID on data files

A5.7. the statistical analysis required for rare event counting.

A6. FILING AND ARCHIVING Understands the need for retrieval of data and records. In respect of archiving and specimen storage, has knowledge of:

A6.1. the requirements for clinical governance

A6.2. the requirements for quality management

A6.3. the requirements for adherence to health and safety protocols

A6.4. the Human Tissue Act 2004.

CEP Flow Cytometry guidance to candidates Nov 10 9 DECLARATION I declare that…………………………………………………..has satisfactorily completed Section A Generic Modules at the level of competence required for the Certificate of Expert Practice in Flow Cytometry as required by the Institute of Biomedical Science.

Signed (supervisor)…………………………………

Date…………………………………

CEP Flow Cytometry guidance to candidates Nov 10 10 SECTION B

SPECIALIST MODULES

Candidates are required to complete one specialist module from this section.

B1. HAEMATOLOGY

B1.1. PRE-ANALYSIS (1) SAMPLE SELECTION Is competent to advise: B1.1.1. clinicians and other healthcare professionals on the appropriateness of samples for flow cytometry in the investigation of haematology malignancies

B1.1.2. on optimal timings for the taking of specimens for flow cytometry. E.g. blood, bone marrow, CSF samples

B1.1.3. on procedures for acquisition of samples and their appropriate and timely transport to the laboratory

B1.1.4. quantity of sample required and is aware of implications of insufficient/sub- optimal sampling.

Is able to:

B1.1.5. demonstrate awareness of sample preservatives and their effect on subsequent sample analysis and result interpretation

B1.1.6. demonstrate awareness of effect of pre-analytical variable on subsequent sample analysis and result interpretation

B1.2. PRE-ANALYSIS (2) SAMPLE PREPARATION

B1.2.1. Has a thorough understanding of the principles of sample preparation including:  health and safety issues relating to the use of hazardous chemicals  health and safety issues associated with infectious materials  hazards associated with inactivation of infectivity  sample quality and quantity.

Is competent to: B1.2.2. prepare reagents of suitable quality for flow cytometry

B1.2.3. prepare samples for examination using procedures in accordance with HPA standard operating procedures, the BCSH task force and the WHO guidelines

B1.2.4. identify other methodologies that may be more appropriate in relevant circumstances and advise clinicians and other healthcare professionals accordingly. Understands the importance of: B1.2.5. clinical information in determining sample processing

B1.2.6. communication with the relevant pathologist.

CEP Flow Cytometry guidance to candidates Nov 10 11 B1.3. CELL LABELLING Has demonstrable experience of: B1.3.1. selecting appropriate reagents to use

B1.3.2. labelling different types of sample

B1.3.3. producing good quality samples for analysis

B1.3.4. assessment of labelling quality

B1.3.5. the range of methods for labelling including direct, indirect and intracellular techniques

B1.3.6. artefacts produced by labelling techniques

B1.3.7. potential problems associated with sample storage.

B1.4. PROCESSING Has knowledge and understanding of: B1.4.1. principles of sample processing, to include factors affecting the practical aspects of the process

B1.4.2. effects on subsequent procedures with particular regard to labelling produced by poor processing

B1.4.3. sample processing schedules appropriate for use with different sample types.

B1.5. PERMEABILISATION Has knowledge and understanding of: B1.5.1. the general principles of permeabilisating cell suspensions to include factors affecting subsequent procedures

B1.5.2. artefacts produced by permeabilisation

B1.5.3. specific types of commonly used reagents and their characteristics, their suitability and compatibility with subsequent labelling procedures

B1.5.4. problems associated with specimen storage.

B1.6. ANALYSIS OF SAMPLES

B1.6.1. Can identify cell types and debris in samples.

B1.6.2. Can identify the usual features of samples commonly encountered within the candidate’s laboratory.

B1.6.3. Recognises artefacts produced by permeabilisation, processing and labelling techniques including autofluorescence and clumping.

B1.6.4. Understands the importance of appropriate labelling and operating parameters of the flow cytometer to the quality of the analysis.

B1.6.5. Is competent at recording data files on the flow cytometer and re-analysing them.

CEP Flow Cytometry guidance to candidates Nov 10 12 B1.6.6. Understands the statistical analysis required for rare event counting.

B1.6.7. demonstrate evidence of participation in a suitable external quality assurance scheme

B1.6.8. demonstrate participation in an internal quality assurance scheme for the use of the flow cytometry in haematology

B1.7. POST-ANALYSIS

B1.7.1. Can describe the implications of presence, increase, decrease or absence of cells by flow cytometry associated with haematology malignancy

B1.7.2. Is able to communicate flow cytometry results to the relevant clinicians or healthcare professionals and advise on the implications of such results.

B1.7.3. Is able to provide evidence of flow cytometry findings for purposes of audit and review.

B1.8. INTERPRETATION

B1.8.1. Can interpret the results of the analysis to define the malignancy.

B1.8.2. Understands the scoring systems used.

B1.8.3. Understands the importance of minimal residual disease detection.

B1.8.4. Understands the need to assess the leukaemic/lymphoma phenotype in association with morphological examination and cytogenetic analysis as appropriate.

DECLARATION I declare that…………………………………………………..has satisfactorily completed the specialist module B1 Haematology at the level of competence required for the Certificate of Expert Practice in Flow Cytometry as required by the Institute of Biomedical Science.

Date…………………………………

CEP Flow Cytometry guidance to candidates Nov 10 13 B2. IMMUNOLOGY

B2.1. PRE- ANALYSIS (1) Is competent to advise: B2.1.1. clinicians and other healthcare professionals on the appropriateness of samples for flow cytometry

B2.1.2. on optimal timings for the taking of specimens for flow cytometry

B2.1.4. on quantity of sample required and is aware of implications of insufficient/sub- optimal sampling. Is able to: B2.1.5. demonstrate awareness of sample preservatives and their effect on subsequent sample analysis and result interpretation

B2.2.1. Has thorough understanding of the principles of sample preparation including:  Health and safety issues relating to the use of hazardous chemicals  Health and safety issues associated with infectious materials  Hazards associated with inactivation of infectivity  Sample quality and quantity. B2.2.2. Is competent to identify samples that need referral to a specialist centre for morphological examination.

B2.2.3. Is competent to prepare reagents of suitable quality for flow cytometry.

B2.2.4. Is competent to prepare samples for examination using the following procedure: lymphocyte markers in accordance with HPA standard operating procedures.

B2.2.5. Is aware and understands the significance of other procedures: HLA Class I and HLA ClassII markers, neutrophil cell adhesion molecules.

B2.2.6. Can identify other methodologies that may be more appropriate in relevant circumstances and advise clinicians and other healthcare professionals accordingly.

B2.2.7. Understands how to prepare samples for the measurement of neutrophil respiratory burst by flow cytometry.

B2.2.8. Is aware of sample preparation for measurement of lymphocyte proliferation.

CEP Flow Cytometry guidance to candidates Nov 10 14 B2.3. ANALYSIS

B2.3.1. Is able to distinguish different cell types.

B2.3.2. Is competent to recognise circumstances when preparation is likely to have affected cell morphology.

B2.3.3. Is competent to identify unexpected cells present in clinical preparations.

B2.3.4. Is able to quantify numbers of different cell types.

B2.3.5. Understands the gating strategy and calculation of absolute lymphocyte counts and how to interpret flow cytometry data.

B2.3.6. Understands the statistical analysis required for rare event counting.

B2.3.7. demonstrate evidence of participation in a suitable external quality assurance scheme

B2.3.8. demonstrate participation in an internal quality assurance scheme for the use of the flow cytometry in immunology

B2.4. POST-ANALYSIS

B2.4.1. Can describe the implications of presence, increase, decrease or absence of cells by flow cytometry.

B2.4.2. Is able to communicate flow cytometry results to the relevant clinicians or healthcare professionals and advise on the implications of such results.

B2.4.3. Is able to provide evidence of flow cytometry findings for purposes of audit and review.

B2.5.1. Identify clinically relevant levels of cells and markers.

B2.5.2. Can interpret neutrophil respiratory burst.

B2.5.3. Is aware how to interpret lymphocyte proliferation data.

DECLARATION I declare that…………………………………………………..has satisfactorily completed the specialist module B2 Immunology at the level of competence required for the Certificate of Expert Practice in Flow Cytometry as required by the Institute of Biomedical Science.

Signed (supervisor) …………………………………

Date …………………………………

CEP Flow Cytometry guidance to candidates Nov 10 15 B3. HISTOCOMPATABILITY AND IMMUNOGENETICS

B3.1. PRE- ANALYSIS (1) Is competent to advise:

B3.1.1. clinicians and other healthcare professionals on the appropriateness of samples for flow cytometry

B3.1.2. on optimal timings for the taking of specimens for flow cytometry, e.g. pre- transplantation and post-transplantation

B3.1.4. on quantity of sample required and is aware of implications of insufficient/sub- optimal sampling.

Is able to demonstrate:

B3.1.5. demonstrate awareness of sample preservatives and their effect on subsequent sample analysis and result interpretation

B3.2.1. Has a thorough understanding of the principles of sample preparation including:  Extract donor lymphocyte targets from spleen or peripheral blood by Ficoll- Hypaque centrifugation prior to flow cytometry crossmatching  The effective removal of platelets from peripheral blood prior to flow cytometry crossmatching  Health and safety issues relating to the use of hazardous chemicals  Health and safety issues associated with infectious materials  Hazards associated with inactivation of infectivity  Sample quality and quantity.

Is competent to:

B3.2.2. identify samples that need referral to a specialist centre for eg HLA-B27 in the diagnosis of ankylosing spondylitis

B3.2.3. prepare reagents of suitable quality for flow cytometry

B3.2.4. prepare samples for examination using the following procedures in accordance with HPA standard operating procedures

B3.2.5. identify other methodologies that may be more appropriate in relevant circumstances and advise clinicians and other healthcare professionals accordingly.

B3.3.1. Is able to identify the cell population by its forward and side scattering properties.

B3.3.2. Can analyse the lymphocytes and generate a dot plot from which the positive stained T and B cells can be identified.

B3.3.3. Can display a histogram of fluorescence (IgG) profile of the T and B lymphocytes to show the degree of IgG binding.

B3.3.7. Demonstrate evidence of participation in a suitable external quality assurance scheme.

B3.3.8. Demonstrate participation in an internal quality assurance scheme for the use of the flow cytometry in histocompatability and immunogenetics

B3.4. POST-ANALYSIS

B3.4.1. Can compare the average of the median T cell and B cell peak FITC fluorescence test sera; calculate the difference between the two and express as a positive or negative result.

Is able to:

B3.4.2. distinguish between mean channel florescence (MCF) and mean channel shift (MCS) when reporting results.

B3.4.3. communicate flow cytometry results to the relevant clinicians or healthcare professionals and advise on the implications of such results.

B3.4.4. provide evidence of flow cytometry findings for purposes of audit and review.

B3.5.1. Can identify which antibodies are clinically important.

B3.5.2. Is aware that the definition of positivity needs to be established in each centre performing the crossmatch.

CEP Flow Cytometry guidance to candidates Nov 10 17 DECLARATION I declare that…………………………………………………..has satisfactorily completed the specialist module B3 Histocompatibility and Immunogenetics at the level of competence required for the Certificate of Expert Practice in Flow Cytometry as required by the Institute of Biomedical Science.

Date …………………………………

CEP Flow Cytometry guidance to candidates Nov 10 18 B4. STEM CELL BIOLOGY

B4.1. PRE- ANALYSIS (1) Is competent to advise: B4.1.1. clinicians and other healthcare professionals on the appropriateness of samples for flow cytometry

B4.1.2. on optimal timings for the taking of specimens for flow cytometry, e.g. pre- transplantation and post- transplantation

B4.1.3. on procedures for acquisition of samples and their appropriate and timely transport to the laboratory, due to the viability maintenance

B4.1.4. on quantity of sample required and be aware of implications of insufficient/sub- optimal sampling. Is able to demonstrate: B4.1.5. awareness of sample preservatives and their effect on subsequent sample analysis and result interpretation

B4.1.6. awareness of effect of pre-analytical variable on subsequent sample analysis and result interpretation

B4.1.7. participation in a suitable external quality assurance scheme

B4.1.8. participation in an internal quality assurance scheme for the use of the flow cytometer in transplantation.

B4.2.1. Has a thorough understanding of the principles of sample preparation including:

 health and safety issues relating to the use of hazardous chemicals

 health and safety issues associated with infectious materials

 hazards associated with inactivation of infectivity

 sample quality and quantity.

B4.2.2. Is competent to prepare reagents of suitable quality for flow cytometry.

B4.2.3. Is aware of different antibody classes used for the analysis.

B4.2.4. Is competent to prepare samples for examination using procedures in accordance with HPA standard operating procedures.

B4.2.5. Is aware, understand and be able to follow the Medicines and Healthcare products Regulatory Agency (MHRA) standards and work in accordance with such standards when obtaining samples in the sterile environment (e.g. in the case of allogenic transplants).

B4.3.6. Demonstrate evidence of participation in a suitable external quality assurance scheme

B4.3.7. Demonstrate participation in an internal quality assurance scheme for the use of the flow cytometry in stem cell biology

B4.4. POST-ANALYSIS

B4.4.1. Can describe the implications of presence or absence of cells by flow cytometry.

Is able to:

B4.4.2. communicate flow cytometry results to the relevant clinicians or healthcare professionals and advise on the implications of such results

B4.5.1. Can identify clinically relevant levels of cells.

DECLARATION I declare that…………………………………………………..has satisfactorily completed the specialist module B4 Stem Cell Biology at the level of competence required for the Certificate of Expert Practice in Flow Cytometry as required by the Institute of Biomedical Science.

Date…………………………………

CEP Flow Cytometry guidance to candidates Nov 10 20 B5. TRANSFUSION

B5.1. PRE-ANALYSIS (1) Is competent to advise: B5.1.1. clinicians and other healthcare professionals on the appropriateness of samples for flow cytometry

B5.1.2. on optimal timings for the taking of specimens for flow cytometry

B5.1.4. on quantity of sample required and be aware of implications of insufficient/sub- optimal sampling Is able to demonstrate awareness of: B5.1.5. sample preservatives and their effect on subsequent sample analysis and result interpretation

B5.1.6. effect of pre-analytical variable on subsequent sample analysis and result interpretation.

B5.2.1. Has a thorough understanding of the principles of sample preparation including:  Health and safety issues relating to the use of hazardous chemicals  Health and safety issues associated with infectious materials  Hazards associated with inactivation of infectivity  Sample quality and quantity. Is competent to: B5.2.2. identify samples that need referral to a specialist centre, e.g. foeto-maternal bleed (Kleihaeur)

B5.2.3. prepare reagents of suitable quality for flow cytometry

B5.2.4. prepare samples for examination using the following procedures in accordance with HPA standard operating procedures.

B5.3. ANALYSIS

B5.3.6. Evidence of participation in a suitable external quality assurance scheme

B5.3.7. Demonstrate participation in an internal quality assurance scheme for the use of the flow cytometer in transfusion.

CEP Flow Cytometry guidance to candidates Nov 10 21 B5.4. POST-ANALYSIS

B5.4.1. Can describe the implications of presence or absence of cells by flow cytometry.

Is able to:

B5.4.2. communicate flow cytometry results to the relevant clinicians or healthcare professionals and advise on the implications of such results

B5.5.1. Can identify samples with clinically significant levels of cells.

DECLARATION I declare that…………………………………………………..has satisfactorily completed the specialist module B5 Transfusion Science at the level of competence required for the Certificate of Expert Practice in Flow Cytometry as required by the Institute of Biomedical Science.

Date …………………………………

CEP Flow Cytometry guidance to candidates Nov 10 22 SUGGESTED READING LIST

JOURNALS 1. Cytometry. The Journal of the International Society for Analytical Cytology 2. Journal of Immunological Methods

BOOKS 1. Givan A L. Flow cytometry first principles. 3rd edn. John Wiley & Sons, 2009. ISBN-10: 047018308X 2. McCarthy DA, Macey MG, eds. Cytometric analysis of cell phenotype and function. Cambridge University Press, 2001. ISBN-10: 0521660297 3. Macey M G ed. Flow cytometry: Principles and Applications. 1st edn. Humana Press, 2007. ISBN-10: 1588296911 4. Ormerod MG ed. Flow cytometry: a practical approach. 3rd rev edn. Oxford University Press, 2001. ISBN-10: 019963825X 5. Robinson J P, ed. Handbook of flow cytometry methods. New York. Wiley-Liss, 1993. ISBN-10: 0471596345 6. Shapiro H M. Practical flow cytometry. 4th rev edn. John Wiley & Sons, 2003. ISBN-10: 0471411256

WEBSITE LINKS www.isac-net.org www.nfcr.lanl.gov www.classimed.de/cytorel.html www.cytometry.org www.cyto.purdue.edu http://pingu.salk.edu/fcm/sitelink.html http://www.meds.com/leukemia/leukemia.html http://www.meds.com/leukemia/flow/flow0.html http://www.hss.edu/11564.asp http://www.ncbi.nlm.nih.gov/ http://www.bork.embl.de/j/ http://www.rms.org.uk/ http://www.immune-source.com/html/favorite_links.html www.dti.gov.uk http://www.antibodyresource.com/

CEP Flow Cytometry guidance to candidates Nov 10 23