TrgnsFj/ CSF Sample Storage Tube ce IVD Safety Data Sheet can be obtained at www.caltag.co.uk H317 - May cause an allergic skin reaction. P261 - Avoid breathing fume, mist, spray, vapours. P280 - Wear protective gloves, protective clothing, eye protection. P302+P352 - IF ON SKIN: Wash with plenty of soap and water. P308+P313 - IF exposed or concerned: Get medical advice/attention. P362+P364 - Take off contaminated clothing and wash it before reuse. INSTRUCTIONS FOR USE Intended Purpose TransFix® Cerebrospinal Fluid (CSF) Sample Storage Tubes are intended for the stabilisation and storage of human CSF specimens for characterisation of infiltrated white blood cells by flow cytometry. Recovery of infiltrated leucocyte subsets can be accomplished over a 3-day period following stabilisation. TransFix CSF Sample Storage Tubes are for In Vitro Diagnostic Use. Within CSF specimens, the qualitative and quantitative characterisation of infiltrated leucocytes via flow cytometry is an important tool for identification of various immunodeficiencies and haematologic diseases [1-4]. However, the number of leucocytes present in CSF is often very low and degrade quickly. Therefore, assessment of these cells is reliant upon urgent analysis which is not always possible. Subsets of leucocytes can be distinguished by cell surface antigens using fluorescent antibodies and flow cytometry. The TransFix stabiliser acts by preserving cell surface antigens of lymphocyte subsets until processing and analysis can be performed. Addition of CSF to TransFix CSF Sample Storage Tubes at the time of lumbar puncture has been shown to significantly extend the integrity of these leucocytes compared to unstabilised samples or stabilisation by other methods [1, see Representative Data Figure 1]. TransFix preserves the cell surface antigens until processing and analysis can be performed [1, 2]. TransFix CSF 1-4 mL Sample Storage Tubes consist of 5 mL polypropylene vials containing 0.2 mL TransFix suitable for stabilising 1-4 mL of CSF. TransFix CSF 0.25-1 mL Sample Storage Tubes consist of 2 mL polypropylene vials containing 0.05 mL TransFix suitable for stabilising 0.25-1 mL of CSF. 1. TransFix CSF Sample Storage Tube is intended for use as specified in this document. It is an 'In Vitro Diagnostic' product for professional use only. 2. TransFix CSF Sample Storage Tubes are only for use with flow cytometry analysis. 3. TransFix CSF Sample Storage Tubes must be stored at 2-8°C. Do not freeze. 4. Do not use after the expiration date on the tubes and packaging. 5. Do not dilute or add other components to the tube before use. 6. Do not use cell viability stains on samples treated with TransFix as they are fixed instantaneously. 7. TransFix CSF Sample Storage Tubes are single-use only. Do not re-use. 8. Ensure the correct sample volume is used as per instructions; an incorrect sample-to-additive ratio may lead to poor product performance and incorrect analytic results. 9. TransFix treated samples and all materials coming into contact with it should be handled as if capable of transmitting infection. TransFix CSF Sample Storage Tubes contain EDTA and TransFix, a clear green liquid containing formaldehyde and other chemicals. Indications of Product Deterioration 1. Cloudiness or precipitate visible in the TransFix. 2. Colour change of TransFix from a clear green liquid. 3. TransFix change from liquid to solid. If indications of product deterioration occur, do not use and contact Caltag immediately on: +44(0)1280 827460 or [email protected]. All TransFix products are shipped at ambient temperature (2 - 25°C). Additional insulation may be required for shipment during extreme temperature conditions. TransFix CSF Sample Storage Tubes must be stored at 2-8°C on arrival. Materials needed but not supplied • Sample collection tubes and materials for sampling. • Automatic pipettes with disposable tips for 10, 20, 100 and 1,000 pL. • Flow cytometry tubes • Flow cytometry calibration and control reagents • Specific fluorescent antibodies and isotype controls • Automatic agitator/vortex machine • Flow cytometer 1. Collect cerebrospinal fluid (CSF) by lumbar puncture according to the relevant clinical standard. 2. TransFix 1-4 mL CSF Tubes: Carefully mix the sample and transfer 1-4 mL CSF into the 5 mL tube using a manual pipette as soon as possible and no later than 1 hour after lumbar puncture. 3. TransFix 0.25-1 mL CSF Tubes: Carefully mix the sample and transfer 0.25-1 mL CSF into the 2 mL tube using a manual pipette as soon as possible and no later than 1 hour after lumbar puncture. 4. Gently mix the TransFix treated sample and close the cap. 5. Store / transport the TransFix treated sample for up to 72 hours at 2-8°C. 6. Stain and analyse your stabilised CSF sample as per your usual CSF flow cytometry protocol (N.B. see Notes) 1. It is recommended that all antibody conjugates are validated in association with TransFix CSF Sample Storage Tubes prior to use. 2. Use caution when implementing automatic gating strategies as light scatter positions of cells stabilised by TransFix may differ from those of untreated cells. 3. Gating for lymphocytes should be performed on CD45+/SSc. 4. If calculating absolute cell counts, the dilution with TransFix must be accounted for. I.e., for a sample size of 1 mL adjust the absolute cell count by multiplying the output by 1.2. For a sample size of 2 mL adjust the absolute cell count by multiplying the output by 1.1, etc. TransFix contains formaldehyde which should be disposed of in accordance with local regulations. Avoid disposing into drainage systems and the environment. Once the TransFix CSF Sample Storage Tube contains a biological sample it must be disposed of in accordance with local regulations regarding clinical waste. 1. Use of TransFix Cerebrospinal Fluid Storage Tubes Prevents Cellular Loss and Enhances Flow Cytometric Detection of Malignant Haematological Cells after 18 Hours of Storage. De Jongste et. al., Cytometry Part 2. Guidelines on the use of Multicolour Flow Cytometry in the Diagnosis of Haematological Neoplasms. Johansson et. al., British Journal of Haematology, 2014, 165, 455-488. 3. Flow Cytometric Characterization of Cerebrospinal Fluid Cells. De Graaf MT et. al., Cytometry, 2011, 80B: 271-281. 4. ESCCA/ISCCA Protocol for the Analysis of Cerebrospinal Fluid by Multiparametric Flow-cytometry in Hematological Malignancies. Del Principe et. al., Cytometry, 2020; 1- 13. Symbols Glossary For symbols glossary, please see https://www.caltag.co.uk/resources/product-information/ Ordering Information Please call Caltag on +44(0)1280 827460 or email [email protected] for assistance. Additional information can be found online at www.caltag.co.uk. A certificate of conformity can be provided with every batch of TransFix CSF Sample Storage Tubes. In case of any serious incident, please contact the legal manufacturer Caltag and the competent authority of the Member State in which the user and/or the patient is established.