Immunofluorescence

Written by Robert Storer.

 

Direct immunofluorescence is used for the detection of tissue bound proteins such as antibodies and complement. Two types of tissue, skin or renal, are processed, although occasionally others may be submitted (e.g. oral mucosa, conjunctiva). Biopsies are taken from patients; snap-frozen and sectioned prior to incubation with fluorescein isothiocyanate (FITC) conjugated anti-sera. This method allows for the detection of tissue bound antibodies when there may be insufficient levels to detect in serum. Other tissue reactants such as complement and fibrinogen can also be detected. In some disease states patterns of staining can be diagnostic.


Instructions for Collection of Samples

 

Do NOT Put Samples For IMF in to Formalin

 

Skin Samples

Skin samples are wrapped in saline soaked gauze and place in a universal container with full patient details label attached.  The skin samples must include the dermal-epidermal junction (DEJ) for tests to be performed.

 

Renal Samples

Renal samples are placed in saline solution in a universal container with full patient details label attached.  Renal samples must include the portion of a kidney containing glomureli.

Any other tissue sample requiring IMF must be placed in saline.  Formalin is NOT an appropriate medium; IMF tests cannot be performed on formalin fixed tissue.

 

 

 

Sending samples to Cellular Pathology department

Samples from patients with known infectious risks ie. Hep B, TB, HIV must be discussed with a pathologist or Immunologist before the sample is taken.

Fill out a Histology form with all the patient and corresponding clinical details. The form must be clearly labelled IMF.

If a sample for routine histology has been taken alongside the IMF sample then one form can be sent detailing the pots and instructions for tests needed.  All samples taken on one patient are to be sent together (thus one uniform report can be generated).

 Samples requiring IMF analysis must be sent as soon as possible, within 24 hours of collection to ensure optimal results.

 

For further information contact

                Mr Gavin Rock – Biomedical Scientist – 0121 424 3188

                Dr Peter Colloby, Dermatology specialist – 0121 424 2921

                Dr Gerald Langman, Renal Specialist – 0121 424 3189

HER 2 Testing Service

Written by Robert Storer.

We perform over 2000 HER2 Immunohistochemistry and 800 FISH tests per year, on breast cancer cases, using FDA approved and validated assays ( Pathway 4B5 antibody from Roche/Ventana and PathVysion dual colour FISH probe from Abbott Molecular (both CE marked)
All testing is performed in accordance with National Guidelines.

References:

  • Bartlett JMS, Starczynski J et al. HER2 testing in the UK: recommendations for breast and gastric in-situ hybridisation methods. J Clin Pathol 2011; 64: 649-653)
  • Woolff AC, Hammond EH et al. American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Human Epidermal Growth Factor 2 Testing in Breast Cancer. J Clin Oncol 2007; 25 (1): 118-145
  • Walker RA, Bartlett JMS et al. HER2 testing in the UK: further update to recommendations. J Clin Pathol 2008; 61: 818-824
  • Ellis IO, Dowsett M et al. Recommendations for HER2 testing in the UK. J Clin Pathol 2000; 53: 890-2
  • Ellis IO, Bartlett J et al. Updated recommendations for HER2 testing in the UK. J Clin Pathol 2004; 57: 233-7)

We are a national reference centre laboratory for HER2 testing
We participate in national UKNEQAS scheme for HER2 testing
We are actively involved in monitoring and auditing all aspects of the HER2 service to best meet the need of our users.

Specimen Labelling And Completion Of The Request Form

Written by Robert Storer.

All Histology specimens must be labelled appropriately and accompanied by a fully completed request from.

Specimen Labelling Requirements

  • Forename and surname
  • Date of birth
  • NHS number and or Patient Identification number
  • Address
  • Nature of specimen
  • Request Form Labelling Requirements

  • Forename and surname
  • Date of birth
  • NHS number and or Patient Identification number
  • Address
  • Nature of specimen
  • Clinical features
  • Any known concomitant diseases
  • Requesting doctor
  • Location
  • Date and time specimen taken
  • Items in bold type are essential. Failure to provide the essential information may result in a delay in reporting as confirmation of patient demographics will be required prior to specimen processing Where there are inadequate patient or specimen identification details the clinician will be informed and requested to attend the department, identify the case and provide the necessary details. If the clinician cannot be contacted the request form and specimen will be returned to the sender with a "discrepancy" proforma to be completed by the clinician. The completed and signed proforma must then be returned with the corrected form/specimen pot to the department.

    If a report is required urgently the request form must be labelled URGENT and ensure the specimen is sent to the laboratory as soon as possible. If a report is required for an MDT it is helpful if the date of the MDT is stated on the form.