Why Stabilise Cerebrospinal Fluid with TransFix?

Cerebrospinal fluid (CSF or liquor) is a colourless bodily fluid found in the brain and spine which
acts as a cushion or buffer from the brain’s cortex. CSF is obtained from the body via lumbar
puncture; the back is anaesthetised and a needle inserted between the 3rd and 4th lumbar
vertebrae. CSF is then collected under its own pressure.

Analysis of white blood cells in CSF is critical in the diagnosis of many infectious and inflammatory
neurological disorders. As well as central nervous system involvement with lymphoproliferative disorders.

Flow cytometric analysis can provide absolute cell counts and detect rare events with high sensitivity
and specificity, however cells within CSF samples are typically low in number, sparse in concentration
and degrade rapidly ex vivo. This results in the necessity to promptly test CSF specimens to prevent
false negative results (1)(2).

Features of TransFix

TransFix is a solution that stabilises leucocytes and leucocytic antigens for up to 14 days in multiple
sample types, preserving the absolute cell counts and cellular markers. This allows for the differentiation
of leucocyte sub-populations and provides an extended time frame for analytical testing.

TransFix has been shown to stabilise malignant haematological cells in cerebrospinal fluid, making it
possible to determine leucocyte subsets in CSF via flow cytometric analysis 72 hours after lumbar puncture. 
TransFix has therefore been has been recommended in guidelines published in the British Journal of
Haematology (see case study 1).

• TransFix/EDTA CSF Sample Storage Tubes are specifically designed for the stabilisation of cells
  within CSF specimens.
• Quality controlled against CSF substitute, including the markers CD14, CD4, CD8, CD19, CD20,
  Igκ, Igλ, CD56. (see CoAs for full details).
• Specific protocol for CSF sample preparation with TransFix (see IFU).
• Higher lymphocyte yield after 30 minutes compared to untreated samples (see case study 2).
• The cellular component of CSF is stabilised for at least 72 hours allowing for determination of
  cell sub-populations and follow up analysis (see case study 3).
• Light scatter and key antigen expression patterns are comparable to fresh samples
  (see case study 3and case study 4).
• Markers stabilised include Markers stabilised include CD10, CD19, CD34
  (see case study 3 and case study 4), CD45, HLA-DR, and CD117 (see case study 3).

Benefits of Cerebrospinal Fluid Stabilisation

• More time in which to conduct testing
• Allows for further investigation after initial cytomorphological analysis has identified malignant cells,
  using an aliquot of the same sample.
• Allows determination of leucocytic cell populations via flow cytometric analysis, up to 3 days after collection.
• Reduced cell lysis during transportation of samples from the collection site to the laboratory.
• Reduced need for repeat lumbar puncture
  • Reduced costs
  • Reduced trauma for child patients and parents
  • Reduced time spent re-sampling
• Standardise Results:
  • The ability to batch samples for transportation and testing
  • Reduces lab variability
  • Beneficial for clinical research studies: 
  • Haematologica 2014;99(7) p1228-123

    TransFix Stabilisation of Malignant Haematological Cells in Cerebrospinal Fluid
    Case Study 1: Guidelines on the use of multicolour flow cytometry in the diagnosis
    of haematological neoplasms.
    Case Study 2: Use of TransFix Cerebrospinal Fluid (CSF) Storage Tubes Prevents
    Cellular Loss and Enhances Flow Cytometric Detection of Malignant Hematological
    Cells After 18 Hours of Storage
    Case Study 3: Infiltration of CNS by acute leukaemia: Analysis of fresh and stabilised CSF
    Case Study 4: CSF Screening for Suspected CNS localised leukemia or lymphoma

  • Product Information

    TransFix/EDTA CSF Sample Storage Tubes consist of 5ml screw cap tubes containing 0.2 ml of
    Transfix/EDTA, which can be used to stabilise up to 5ml of CSF. These tubes are available in
    packs of 10, 25 and 50. TransFix/EDTA CSF Sample Storage Tubes must be stored at 2-8°C.

    

    Product Code	Description	Quantity
    TF-CSF-5-2	Storage tubes containing 0.2ml TransFix/EDTA	2 x 5ml
    TF-CSF-5-10	Storage tubes containing 0.2ml TransFix/EDTA	10 x 5ml
    TF-CSF-5-25	Storage tubes containing 0.2ml TransFix/EDTA	25 x 5ml
    TF-CSF-5-50	Storage tubes containing 0.2ml TransFix/EDTA	50 x 5ml

    References

    1. de Graaf, M.T. et al. (2011). Flow Cytometric Characterization of Cerebrospinal Fluid Cells.
       Cytometry Part B [online] Volume 80B(5), p. 271–281.
       Available at: http://onlinelibrary.wiley.com/doi/10.1002/cyto.b.20603/abstract [Accessed 11 Nov. 2016].
    2. Kraan, J. et al (2008) Flow cytometric immunophenotyping of cerebrospinal fluid.
       Current Protocols in Cytometry [online] Volume 45, 6.25.1–6.25.16.
       Available at: http://onlinelibrary.wiley.com/doi/10.1002/0471142956.cy0625s45/abstract [Accessed 11 Nov. 2016].
    3. Johansson U, Bloxham D, Couzens S, Jesson J, Morilla R, Erber W, Macey M and British Committee for Standards in Haematology.
       (2014) Guidelines on the use of multicolour flow cytometry in the diagnosis of haematological neoplasms.
       British Journal of Haematology 165: p. 455-488.
       Available at: http://onlinelibrary.wiley.com/doi/10.1111/bjh.12789/full [Accessed 20 Nov. 2016]
    4. Brem, S.S. et al. (2008) NCCN clinical practice guidelines in oncology.
       Central nervous system cancers.
       V. 1.2008 ed: http://www.nccn.org/professionals/physician_gls/PDF/cns.pdf [Accessed 11 Nov. 2016].