Dubowitz Neuromuscular Centre
Hammersmith Hospitals NHS Trust
726968980
Page 1 of 4
Imperial College Referral Centre for Congenital Muscular Dystrophies Edition: 1.0
Funded through the National Specialist Commissioning Advisory Group (NSCAG). Department of Health
Section 1: General Information
Send blood & DNA samples and a copy of the corresponding referral form to:
Mrs Rachael Mein. Genetics Centre
8 th Floor Guy’s Tower. Guy’s Hospital
London SE1 9RT. Tel: 020 718 81714.
Send all other samples and the original of all referral forms to:
Dr. Lucy Feng
NSCAG Referral Centre for Congenital Muscular Dystrophy
Dubowitz Neuromuscular Centre. Hammersmith Hospital
Du Cane Road. London W12 ONN
Please enclose a completed form and any relevant information including a biopsy report if available
All specimens must be sent by prior arrangement by contacting:
Dr.Lucy Feng: l.feng@ic.ac.uk
; Ms Jihee Kim: jhkim@imperial.ac.uk
; Dr. Cecilia Jimenez-Mallebrera: c.jimenez@ic.ac.uk
on Tel: 020 8383 2126; Fax: 020 8746 2187. Alternatively contact Ms Phil Davies
(Administrator): pmdavies@hhnt.nhs.uk
on Tel: 020 8383 2127.
Write the extension number of the lab (32126) on top of the parcel
Do not send samples for delivery on a Friday or Saturday. The best days to send samples are Monday to Wednesday. The Laboratory opening hours are 9 AM- 5:30 PM
Samples/containers must be clearly labelled with the patient name and date of birth or another second identifier (e.g. hospital number, laboratory specimen number)
** NOTE: We are not able to accept samples designated, or suspected, as being "High Risk" (i.e. from patients with HIV, hepatitis, polio, CJD etc)
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AMPLES THAT ARE NOT SENT ACCORDING TO THESE INSTRUCTIONS MAY NOT BE SUITABLE FOR
ANALYSIS
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Section 2: Types of samples that we accept
Muscle and or skin biopsy, chorionic villi, blood, DNA , frozen muscle, OCT block and fibroblast culture, histological slides for second opinion
Section 3: Handling of samples
3.1 Samples from Hammersmith Hospital
Specimens should be brought to the lab (as soon as possible after being obtained) in the following containers/media
Muscle biopsy
Collect the biopsy in a sterile gauze dampened in sterile saline and place inside a screw cap 30 ml universal (or other safely sealed) sterile container
When obtaining an open muscle biopsy (e.g for respiratory chain enzymes analysis) collect a piece of at least 1 cm long x 0.5 cm wide.
Skin biopsy
Immerse the biopsy in approximately 4 ml of sterile standard culture medium (e.g. Dulbecco’s modified
Eagle’s medium) inside a small universal with screw cap.
C.V.S.
Immerse the C.V.S. (5-10 branches) in approximately 4 ml of sterile standard culture medium inside a small 6 ml universal with screw cap.
Blood
Collect in EDTA tube labelled with patient’s name and hospital number
DNA resupended in 1x TE buffer or water for molecular biology- indicate patient’s name, hospital number and DNA concentration in the tube
3.2 Other Hospitals
3.2.1 Duration of sample journey 0-1 hrs
Muscle biopsies must arrive within 1hr of being obtained. If this is not possible proceed as in section 3.2.2. Skin, CVS , blood and DNA samples can withstand room temperatures for a few hours
Muscle biopsy
Collect the biopsy in a sterile gauze dampened in sterile saline and place inside a screw cap 30 ml universal (or other safely sealed) sterile container. DO NOT immerse the biopsy in saline. Send at room temperature.
When obtaining an open muscle biopsy (e.g for respiratory chain enzymes analysis) collect a piece of at least 1 cm long x 0.5 cm wide.

Dubowitz Neuromuscular Centre
Hammersmith Hospitals NHS Trust
726968980
Page 3 of 4
Imperial College Referral Centre for Congenital Muscular Dystrophies Edition: 1.0
Funded through the National Specialist Commissioning Advisory Group (NSCAG). Department of Health
Skin biopsy
In an aseptic manner obtain a piece of skin deep enough to include the dermal layer. Deposit the biopsy in a sterile container (e.g. small universal with screw cap) containing approximately 4 ml of sterile standard culture medium (e.g. Dulbecco’s modified Eagle’s medium without either antibiotics or serum).
Send at room temperature.
C.V.S.
Immerse the clean C.V.S. (5-10 branches) in approximately 4 ml of sterile standard culture medium (as above) inside a small 6 ml universal with screw cap. Send at room temperature.
Blood
Collect in EDTA tube labelled with patient’s name and hospital number. Send at room temperature.
DNA
In 1x TE buffer - indicate patient’s name, hospital number and DNA concentration in the tube. Send at room temperature
Fibroblasts in culture
Send at room temperature 50-100 ml in a flask filled to the top with medium supplemented with 20% serum and sealed. Indicate passage number in the flask.
3.2.2 Duration of sample journey >1hrs
Samples need to be partially processed before being sent to ensure optimum integrity during transport and delivery. Samples have to be processed in a manner adequate for each of the diagnostic tests that will be carried out here in the lab.
Muscle biopsy: Orientate under the dissecting microscope a piece of 30-50mg so as to obtain transverse sections for histology and immuno-histochemistry. Embed in OCT (Tissue-Tek) and freeze in isopentane cooled in liquid nitrogen. For Western blotting, cut a piece of approximately 50 mg from the biopsy. Place inside a cryotube or wrap in a piece of foil and snap freeze in liquid nitrogen. Send the frozen muscle in abundant (2-3Kg) of dry-ice (solid CO
2
).
Skin biopsy, C.V.S., blood, DNA and cells in culture: Proceed as in section 3.2.1
Sample for electron microscopy: Cut sample in small pieces longitudinally. Fix in 4% Glutaraldehyde in
0 .1M Sodium Cacodylate buffer for 1.5 – 2 hours at room temperature (preferably in a glass bijoux).
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Discard the 4% Glutaraldehyde- 0.1M Cacodylate and replace with 0.1M Cacodylate. Leave for 30 min.
After this discard old 0.1M cacodylate and add again 0.1 M Cacodylate. At this point the sample can be stored at 4  C and sent to us at room temperature.
Fibroblasts in culture
Send at room temperature 50-100 ml in a flask filled to the top with medium supplemented with 20% serum and sealed. Indicate passage number in the flask. Alternatively send us a frozen vial in dry-ice.
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ONTACT US FOR ADDITIONAL INFORMATION FOR SAMPLE HANDLING AND FOR UNUSUAL
CIRCUMSTANCES NOT ADDRESSED HERE