Cellular Antigen Stabilising Reagent

Protocol

Product Description

TransFix^® is a patented whole blood stabilisation solution developed by Sheffield Teaching Hospitals NHS Foundation Trust and distributed under licence by Cytomark Ltd.

Intended Use and Principles of Stabilisation

TransFix^® is intended for addition to a previously collected aliquot of blood at a predetermined ratio to act as a stabiliser. The active components of TransFix^® will stabilise cellular antigens for up to 10 days.  In whole blood immunophenotyping TransFix^® is added at a ratio of 200µl per ml (1:5) to K3EDTA anticoagulated blood.  The preparation is then treated as a "routine" sample for immunophenotyping procedures and will maintain its staining profile for up to 10 days when stored at 4°C.

TransFix^® is for in vitro laboratory use and must only be used by individuals trained in the safe handling of potentially infected blood samples.

Products

TransFix^® Single Use Blood Sample Tubes

TransFix^® Bulk

For prices and ordering please contact your local distributor.

Statement of Warnings

1. TransFix^® treated specimens, samples and all material coming in contact with them should be handled as if capable of transmitting infection and disposed of with proper precautions.
2. TransFix^® should be mixed with blood samples within 6 hours of venepuncture. Samples awaiting addition of TransFix^® should be stored at 18-25°C. Do not refrigerate.
3. Avoid contact of TransFix^® and TransFix^® treated samples with skin and mucous membranes. If contact occurs immediately wash off with cold water.
4. Do not use the reagent beyond the expiration date printed on the bottle.
5. Incubation times or temperatures other than those specified may give erroneous results.
6. TransFix^® does not contain anti microbial reagents. Microbial contamination must be avoided or erroneous results may occur.
7. Use Good Laboratory Practices (GLP) when handling this reagent.

Storage and Stability

Store TransFix^® at room temperature (18-25°C). Contents of unopened bottles are stable until the expiration date printed on the label. Contents of opened bottles are stable for 1 month after opening.

Evidence of Deterioration.

The normal appearance of TransFix^® is that of a clear pale green liquid. Any change in the physical appearance of the reagent may indicate deterioration and the reagent should not be used.

Materials Required but Not Supplied

1. Evacuated blood collection tubes with anticoagulant (K3EDTA, ACD, Heparin).
2. Pipettors and plastic tips capable of delivering 200µl to 1000µl.
3. Plastic storage rack.
4. Labels to indicate samples contain TransFix^® reagent.

Anticoagulant

In whole blood samples an EDTA salt (K3EDTA) is the anticoagulant of choice for TransFix^®. However ACD or Heparin may also be used.

Procedure

No preparation is needed. Use TransFix^® reagent directly from the bottle.

1. Determine volume of blood in the blood collection tube. Note: In the Vacuutainer system a full blood collection tube will contain 4ml of blood.
2. Carefully remove blood collection tube cap.
3. Pipette into the blood collection tube, an appropriate volume of TransFix^® at the specified ratio of 200μl TransFix^® per 1000μl of blood.
4. Replace blood collection tube cap, ensuring no leakage is evident.
5. Hand mix by repeated inversion to distribute the TransFix^® reagent completely through the blood sample.
6. Label blood collection tube to indicate that TransFix^® reagent has been added.
7. Place sample in a rack for subsequent cellular antigen assay. TransFix^® treated samples are stable for up to 10 days at 4ºC, 7 days at Room Temperature (18ºC to 24ºC) and for 3 days at 37ºC .

Please note:

Samples stored at 4ºC must be allowed to return to ambient temperature before any cellular antigen assays are performed.

Stored TransFix^® treated samples must be mixed thoroughly by inversion (25x) to resuspend cells before analysis by flow cytometry.

Cell counts from TransFix^® samples must be divided by 0.8 to adjust for the dilution factor.

Limitations

When using TransFix^® with samples collected from leukaemic patients it may be necessary to establish an appropriate volume of reagent to add, which may be different to that quoted in the procedure.  Volumes in the region of 0.5-1.0ml per ml of blood may be more appropriate.