Electron Microscopy Service - specific requirements

The Electron Microscope Unit is used routinely at magnifications of between 1500 and 70,000 times to examine the ultrastructure of cells and their surroundings.  The EM unit receives approx. 800 samples a year, the majority of which are renal, but about 100 muscle and 30 nerve biopsies are also collected.  Other specimens include cardiac biopsies, occasional skin biopsies and tumour samples.

Specimens from outside the department requiring EM should be sent in a small sample container (e.g. bijou or Eppendorf tube) containing at least 2 mL of an EM fixative.  A glutaraldehyde based fixative such as 2.5% glutaraldehyde in phosphate buffer is ideal but 10% buffered formalin is also acceptable.  In larger tissue samples individual pieces should be no larger than 3 mm cubed and should be completely submersed in fixative.  The optimal size of diagnostic tissue requiring processing for electron microscopy is as follows: the largest dimension should be no greater than 5mm and the smallest dimension no less than 1mm. Any specimen with the smallest dimension less than 1mm may not survive tissue processing.

For QEHB service users - Renal biopsies are delivered to the Cellular Pathology Specimen Reception and EM samples are taken in the EM Lab. 

For External service users - package specimens for EM in firmly closed leak proof containers, together with sufficient absorbent material to contain any liquid, in sealed bags. The sealed bags are placed securely in a small padded envelope with the address label as given above under key information.  The EM unit makes use of the Histology request form for both internal and external users.   A copy of the light microscopy report is also required.  

Please note that we support the service by outsourcing when required to the specialist EM facility at Leicester Royal Infirmary.  It is fully accredited by United Kingdom Accreditation Services (UKAS) ISO 15189, reference number 8608.

Advice on the service or requirements can be obtained from the EM lead biomedical scientist This email address is being protected from spambots. You need JavaScript enabled to view it. or from the electron microscopy laboratory team on 0121 371 5720.

Cytopathology

The Department provides the diagnostic cytopathology services for the Trust including fine needle aspirations, endoscopic brushings and washings, EUS-FNAs , serous fluids urines and CSF.  Routine specimens should reach the laboratory before 16:30 so that they can be processed within normal working hours. Specimens collected out of laboratory hours should be stored refrigerated until the next morning on which the laboratory is open.  Cytology sample that do not meet the minimum data requirements on request form or sample, are processed, to ensure that sample integrate is maintained. Details of missing data are added to the report.  Request for Consumables and Request Forms

Sample containers for use in cytology and request forms can be order by contacting the Cytology Department; they can be collected from the Cellular Pathology reception at Heatlands or QEHB sites.  Cytology samples which do not meet the MDS will not be returned.  They will be processed in order to preserve the diagnostic material while attempts are made to contact the sender.  These samples will not be reported until adequate labelling of specimen and request form has been undertaken.Minimal delay between collection and receipt by the laboratory is necessary in order to prevent degeneration of cellular components and consequent loss of diagnostic value. All cytological specimens are potentially biohazardous and must be transported in leak proof containers enclosed in sealed bags. Specimens sent to the laboratory via the SDS should be placed in specimen bags and lids on the all containers should be securely closed. 

Specimens from patients with known or suspected tuberculosis, HIV, viral hepatitis or other transmissible disease should be labelled clearly.  Cytology DO NOT except or handle any specimens that are ? prion disease or CJD.

Specimens should be received as either pre-prepared slides, fresh unfixed samples in white- topped universal containers, or suspended in CRR.  Equipment used to collect the specimen such as brushes or drains are inappropriate.

Sputum - A series of three early morning ‘deep cough’ specimens should be collected on three consecutive days for maximum sensitivity. Post physiotherapy and post bronchoscopy specimens are suitable but should be clearly identified as such.

Serous Fluids - 50mls or the whole volume if less is aspirated should be sent in sterile universal containers. Include any tissue fragments or clots. Do not add fixative of anticoagulant. Drain bags are not suitable for transporting specimens and should NOT be sent.

Cerebrospinal Fluid (CSF) - Rapid processing is essential to preserve cells in CSF. These specimens should be sent to the laboratory within one hour.  ALL CSF SPECIMENS SHOULD REACH THE LABORATORY BY 16:30 AT THE LATEST TO ALLOW PREPARATION.

Urine - A representative aliquot of a maximum of 25ml of urine is sufficient for cytology processing, Ensure that this is not the first sample of the day.  Catheter urine and bladder washings are also acceptable but please mention this on the request form. Mid-stream urine samples are not suitable for cytology because they contain few cells.

Endoscopic Brushing - Immediate fixation is important and all slides must be labelled with PENCIL with the patient’s name and hospital number before the smears are made.

Fine needle aspiration - Unless the clinician is experienced in this procedure, including making good quality smears, it is recommended that they contact the laboratory for advice before beginning the procedure. Smears should be made and the needle rinsed out in cytorich red fluid. For ENT specimens in particular it is important that smears are sent as well as needle washings. Only prepared smears and needle washings are accepted. The sending of needles sheathed or otherwise is strictly forbidden.

EUS-FNAs - Please send all the specimen in cytorich red fluid

Cervical Smears - Cervical smears are NOT accepted in nor prepared by cytopathology at QEHB. Please send any cervical smear specimens to the Cytopathology Laboratory at Wolverhampton's Black Country Partnership Laboratories.

Andrology

The laboratory's priority is to provide a comprehensive semen analysis service for Consultants, Urologists and General Practitioners from the Birmingham and Solihull area. This includes semen analysis (Diagnostic Semen Analysis), post-operative analysis of semen following a vasectomy operation (Post Vasectomy Semen Analysis) and Retrograde analysis of urine.

The laboratory operates on a clinic based service and has a maximum capacity of 2000 appointments available per annum. This covers the annual workload of routine diagnostic semen analysis, post-vasectomy samples and retrograde ejaculation analysis. Routinely patients will be given an appointment to attend the Andrology Department for on-site sample production in a designated private clinic room, however in exceptional circumstances samples may be produced off site if then able to be delivered to the laboratory within an appropriate time interval. 

The UHB Andrology Laboratory has fully trained scientists who are highly proficient in performing quality diagnostic semen analysis in line with World Health Organisation (2021) and the 2016 Laboratory Guidelines for post vasectomy semen analysis. The laboratory regularly performs internal quality control, participates in the UK National External Quality Assurance Reproductive scheme for Andrology (UKNEQAS) and liaises with other accredited laboratories for comparison of tests not covered within the UKNEQAS scope.

A laboratory referral can be made via:

Paper copy referral forms sent to Appointment Centre via post/e-mail

Electronic Referral Service (e-RS) with an attached referral form 

A copy of the referral form can be obtained by;

Contacting the laboratory directly on 0121 424 9717 Or by downloading from here Andrology Referral Form Or by GP’s accessing the word version at: https://www.uhb.nhs.uk/gps/referrals/andrology.htm

Off-site production can be arranged by the patient by telephoning the laboratory on 0121 424 9717 or by e-mailing This email address is being protected from spambots. You need JavaScript enabled to view it. (if you e-mail you are accepting that the connection is not secure and potentially not confidential – laboratory staff will minimise data transmission but cannot guarantee security). Please note that retrograde urine production must be on-site at Good Hope Hospital.

 Turnaround Time: 7-10 days (electronic reporting may be more rapid). Paper copies sent out following result authorisation.

Anti-sperm antibodies (ASAB) are not tested at this laboratory. Agglutination is described if noted.

Please see user handbook for additional information including contact details.

Accreditation information can be found under Cellular Pathology (UKAS 10141): https://www.ukas.com/download-schedule/10141/Medical/

 Post-vasectomy Semen Analysis (PVSA)

The laboratory undertakes examination procedures in line with the following guidelines: 2016 PVSA Guidelines. This will give clinicians information regarding clearance criteria. 

 Results are available to Hospital clinicians via PICS, Clinical Portal, and ICE.

 GP’s can access patient results through their own electronic result systems.

 Key details of the service:

Operates Monday to Friday 08:00am to 16:00pm
Undertakes Diagnostic semen analysis, Post vasectomy semen analysis and Retrograde examination (urine)
Appointment based with on-site private room for sample production
7-10 day turnaround time (electronic and paper copy reporting)
Patients are booked within 7 weeks from referral, unless clinically indicated or by patient choice
The User Handbook can be accessed here: Andrology User Handbook
 

Andrology - Measurement Uncertainty

Following on from our recent UKAS assessment, we were advised that we need to demonstrate that the clinical impact of the Measurement of Uncertainty on clinical decision values has been assessed and considered in relation to patient outcome. This must include an explanation of any effects this could have to the decision limits and how they should be interpreted.

Application of the CV to the clinical relevance of Diagnostic Semen Analysis (DSA)

The Coefficient of Variation (CV) was calculated for each parameter for DSA and used as the measurement of uncertainty associated with each parameter. Each parameter’s CV is as follows:

 The uncertainty is calculated by dividing the analysis result by 100 and multiplying by the associated CV. The range is calculated by +/- the uncertainty from the result.

 The uncertainty could influence the fertility pathway for patients who have received results close to the clinical decision values for each specific parameter. Examples include:

Motility – Uncertainty = 2.2%

This informs us that if the laboratory reports a motility result of 30% then it will have a range of 30% ± 2.2 (27.8 – 32.2%).

Concentration – Uncertainty = 1.4 x 10⁶/ml

This informs us that if the laboratory reports a concentration result of 16 x 10⁶/ml then it will have a range of 16 x 10⁶/ml ± 1.4 (14.6 – 17.4 x 10⁶/ml).

Morphology – Uncertainty = 0.7%

This informs us that if the laboratory reports a morphology result of 4% then it will have a range of 4% ± 0.7 (3.3 – 4.7%).

Vitality – Uncertainty = 1.2%

This informs us that if the laboratory reports a vitality result of 54% then it will have a range of 54% ± 1.2 (52.8 – 55.2%).

Clinical impact of measurement of uncertainty will be communicated to users with annual website updates. Internal users of the service will be emailed directly (fertility and urology).

Clinical Impact of Measurement Uncertainty

Measurement uncertainty, either for one reported parameter or as a combination of multiple reported parameters could have a clinical impact. For example, if a concentration of 15 x 10⁶/ml is reported (oligozoospermia), given a UoM of 1.4 x 10⁶/ml, this sample could have a concentration of 16 x 10⁶/ml (normozoospermia).

For some users, a diagnosis of oligozoospermia, even at the borderline level of 15 x 10⁶/ml, could lead to the recommendation of an assisted reproductive technology such as intracytoplasmic sperm injection (ICSI) to promote the chance of fertilisation, when conventional IVF, or even natural conception, might be possible. 

We inform our users of this uncertainty and emphasize that the reports can be used to guide clinicians to determine how to proceed with further investigation and management of a subfertile couple. We stress that all reported parameters should be considered holistically.

Furthermore, as per WHO 2021 guidance, we inform users that many known and unknown female factors hamper the value of using only semen examination parameters to predict the prognosis for the couple of spontaneous or assisted fertilization.

The decision limits that we provide on our reports are derived from WHO 2021 and are derived from data characterizing the semen characteristics of a reference population (men whose partners had a time to pregnancy of 12 months or less). The data uses a diverse international populations, noting that the percentiles used do not represent distinct limits between fertile and subfertile men.

We highlight that a semen analysis is never prognostic of fertility, as it is the fertility potential of the couple that defines them as fertile or subfertile.

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