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Cytomark

Welcome to Cytomark - A division of Caltag Medsystems

Cytomark are dedicated to the development of cell stabilisation reagents for use in flow cytometry. Based in Buckingham, Cytomark specialise in the development of two key product areas: TransFix® and Blood Controls.

TransFix is a CE marked solution that preserves leucocytes and leucocytic antigens in human peripheral blood for flow cytometric analysis. TransFix is used to extend the timeframe for sample analysis, providing flexibility in testing. Samples treated with TransFix are stabilised for up to 10 days when stored at 2-25°C, and for 4 days at temperatures between 25-37°C. TransFix is widely used for immunophenotyping purposes in hospitals and reference laboratories, as well as academic research institutions, pharmaceutical & biotechnology companies. TransFix can also be used with other cellular samples for research applications.

Cytomark's Blood Controls are reference samples that can be used during the set up and running of flow cytometers to provide quality assurance that equipment & processing procedures are performing optimally. These controls are produced from stabilised human whole blood with both normal and low CD4 counts. Each reference is provided with a QC certificate detailing the expected count and range for T cells (CD3+), T helper cells (CD3+CD4+), T suppressor cells (CD3+CD8+), NK cells (CD56+)and B cells (CD19+).

TransFix - a Cellular Antigen Stabilisation Reagent

TransFix is a CE marked cellular antigen stabilisation reagent that preserves leucocytes and leucocytic antigens in human peripheral blood for flow cytometric analysis. Samples treated with TransFix are stabilised for up to 10 days when stored at 2-25°C, and for 4 days at temperatures between 25-37°C. As a result, samples treated with TransFix® can be analysed at a more convenient time for the operator.

The intended use of TransFix is for immunophenotyping; when added to samples at the point of collection, TransFix immediately stabilises leukocytes for up to 10 days, providing flexibility in analysis. For human whole blood samples, Cytomark routinely QC test that the immunophenotypic profile is maintained for 10 days for cells with markers: CD3, CD4, CD8, CD16, CD19, CD45, and CD56.

Stabilisation of cerebrospinal fluid is also possible using Cytomark's TransFix/EDTA CSF Sample Storage Tubes, which have been developed in collaboration with oncologists specifically for this purpose. Independent research has also been published on the stabilisation of bone marrow samples using the TransFix reagent.

Features and Benefits of TransFix:

  • Stabilises cell surface antigens for a minimum of 10 days at 2-25°C and for 4 days at 37°C
  • Easy to use, just add TransFix and mix by inversion
  • Ensures sample integrity during transportation between clinical sites
  • Eliminates the need for weekend and evening work
  • Reduces the impact of unexpected machine breakdown or staff shortages
  • Allows for repeat testing of samples without the need for patient recall and repeat phlebotomy
  • Greater efficiency in testing - allows for batching of samples prior to testing
  • Reduction in costs associated with the above advantages

Comparison of the leucocytic immunophenotype in normal human whole blood analysed immediately after collection, untreated after 11 days and stabilised with TransFix for 11 days:

Human Blood Stabilisation - Immunophenotyping

Fig. 1 Diagrams A) and B) are flow cytometry density dot plots that demonstrate the normal leucocytic immunophenotype in a fresh sample of human whole blood. Granulocyte, monocyte and lymphocyte populations are all distinct (A) as are T Cells, NK Cells and B Cells (B). Diagrams C) and D) demonstrate the leucocytic immunophenotype in the same sample of human whole blood after 11 days of storage. leucocyte subsets, particularly monocytes are less distinct (C) and B Cells and NK Cells cannot be easily identified (D). Diagrams E) and F) demonstrate the leucocytic immunophenotype in the same sample of human whole blood after treatment with TransFix and storage for 11 days. All leucocyte subsets, including monocytes, B Cells and NK Cells can be clearly identified (E & F).

These images are taken from Cytomark's in-house Quality Control Tests (Cytomark, Buckingham, UK)

References:

Canonico et al (2010). Flow Cytometric Profiles, Biomolecular and Morphological Aspects of Transfixed leucocytes and Red Cells. Clinical Cytometry 78: 267-278

Canonico et al (2004). Evaluation of leuko-cyte stabilization in TransFix-treated blood samples by Flow cytometry and transmission electron microscopy. J Immunol Methods2004;295:67-78.

Barnett et al (1999): TransFix: A clinical sample stabilising fluid for use in clinical haematology and immunology. Cytometry 38:88 (abstract).

To view the full list of publications referencing the use of TransFix on human blood samples, please click here to view the Cytomark Bibliography. If you would like to receive further information about TransFix, please contact us we will be happy to help:

Applications of TransFix: Human Blood Stablilisation

Blood cells begin to degrade as soon as a peripheral blood sample is collected via venepuncture. In fact, blood samples older than 48 hours are no longer suitable for flow cytometric examination. Addition of TransFix to blood samples at the time of collection has been shown to significantly extend the integrity of cells within these samples.

TransFix stabilised samples can be stored and transported at an ambient temperature, removing the requirement for cold chain transportation. This can be a major benefit for outlying clinics and Clinical Research Organisations where samples are routinely transported over long distances.

Clinical Application: Immunophenotyping Human Whole Blood

Blood samples treated with TransFix are suitable for flow cytometric testing, immune monitoring and immunohaematology screening purposes. Samples treated with TransFix are stabilised for up to 10 days when stored at 2-25°C, and for 4 days at temperatures between 25-37°C. Cytomark have demonstrated that the immunophenotypic profile is maintained for up to 10 days, including markers: CD3, CD4, CD8, CD10, CD16, CD19, CD34, CD45, CD56, and cytoplasmic MPO.

Clinical Application: Immune Monitoring of HIV/AIDS

Monitoring CD4 and CD8 T-cell subsets of human immunodeficiency virus (HIV)-infected individuals is performed routinely by flow cytometry. It is the most commonly used method for monitoring the immune status of HIV-infected individuals. Ordinarily, specimens are processed within 36 hours to obtain reliable CD4+ T-cell absolute counts.

Blood treated with TransFix however, remains stable for a minimum of 10 days when stored at 2-25°C and for 4 days at 37°C (1). TransFix stabilises cell surface antigens, allowing transport back to a laboratory without deterioration.

References:

Canonico et al (2010). Flow Cytometric Profiles, Biomolecular and Morphological Aspects of Transfixed leucocytes and Red Cells. Clinical Cytometry 78: 267-278

Canonico et al (2004) Evaluation of leuko-cyte stabilization in TransFix-treated blood samples by Flow cytometry and transmission electron microscopy. J Immunol Methods2004;295:67-78.

Barnett et al (1999): TransFix: A clinical sample stabilising fluid for use in clinical haematology and immunology. Cytometry 38:88 (abstract).

To view the full list of publications referencing the use of TransFix on human blood samples, please click here to view the Cytomark Bibliography. If you would like to receive further information about TransFix, please contact us we will be happy to help:

Tel: 01280 827460
Email: cytomark@caltagmedsystems.co.uk

Applications of TransFix: Stabilisation of Cerebrospinal Fluid

Cells within Cerebrospinal fluid (CSF) samples are typically low in numbers, sparse in concentration and degrade quickly, resulting in the necessity to promptly test CSF specimens to prevent false negative results.

  • TransFix/EDTA CSF Sample Storage Tubes have been developed in collaboration with research oncologists which are optimised for the stabilisation of cerebrospinal fluid.
  • TransFix/EDTA CSF Sample Storage Tubes stabilises leucocytes and leucocytic antigens in CSF for up to 10 days.

References:

Johansson et al (2014). Guidelines on the use of multicolour flow cytometry in the diagnosis of haematological neoplasms. British Journal of Haematology (2014) 165: 455-488

Poster: Infiltration of CNS by acute leukaemia: Analysis of fresh and stabilised CSF - In collaboration with Haematology Dept, Bristol Royal Infirmary, University Hospitals Bristol NHS Foundation Trust, UK. View Here

*Please note that currently TransFix/EDTA CSF Sample Storage Tubes are validated for research use only. They are not validated for immunophenotyping and leukaemia screening. This product is currently undergoing testing with the aim to release as a CE marked in vitro diagnostic medical device.

To find out more about the use of TransFix for CSF stabilisation, including case studies, please click here. Alternatively, please contact us at cytomark@caltagmedsystems.co.uk, or call 01280 827460 for further information.

TransFix: Applications

TransFix can also be used for a number of applications. Find out more about these applications on the Cytomark website by clicking on the links below:

TransFix - Available Formats

TransFix/EDTA Vacuum Blood Collection Tubes (CE)
- Direct draw collection tubes containing TransFix & EDTA to stabilise venous blood at the point of collection

TransFix/EDTA CSF Sample Storage Tubes (RUO)
- In collaboration with leading research oncologists, Cytomark have developed TransFix/EDTA CSF Sample Storage Tubes that are optimised for the stabilisation of cerebrospinal fluid. View our poster here

TransFix Reagent (CE)
- TransFix is provided in 1 and 20ml aliquot sizes and in pack sizes of 10 and 50 1ml tubes. This gives the end user most flexibility with regard to application of TransFix to research samples.

TransFix Sample Storage Tubes (CE)
- 1.2ml tubes containing 0.2ml TransFix (no anti-coagulant)

Custom Sizes
- Cytomark are able to provide TransFix in alternative volumes and quantities. Please contact us with your requirements for further information.

Blood Controls

Routine haematological testing requires the use of reference material during machine set up such that validation prior to analysis of patient samples can be performed. Many also require repeat use of reference controls throughout the running period of the machine, ensuring sample processing, equipment and operators are all performing optimally, thereby providing quality assurance.

In Flow Cytometry however, it has not been the norm to use reference material since it has not previously been possible to stabilise samples for use as a reference. Using proprietary blood stabilisation technology, Cytomark can now produce stabilised human whole blood with both normal and low CD4 counts for use as blood controls. The stabilisation process is robust resulting in controls that are stable for up to 90 days from the date of manufacture when stored at 2 - 8°C, and an open vial stability of 1 month.

The stabilised controls allow gating, and forward and side scatter flow cytometric effects to be determined during machine set up and before and during analysis of patient samples.

The ISO standard entitled ‘ISO 15189: Medical laboratories - Particular requirements for quality and competence’, refers to the requirement for reference samples during flow cytometer set up and running. Cytomark's Stabilised Human Whole Blood and CD4 Low and Normal Controls provide suitable reference samples for this purpose.

Key Benefits of Cytomark's Blood Controls:

  • Verification of monoclonal antibodies and flow cytometry reagents
  • Validation of flow cytometer performance
  • Validation of operator technique
  • Verification of red blood cell lysis

Blood Controls Available:

Stabilised Human Whole Blood

The Stabilised Human Whole Blood control is supplied with a QC certificate detailing the expected count and range for T cells (CD3+), T helper cells (CD3+CD4+), T suppressor cells (CD3+CD8+), NK cells (CD56+)and B cells (CD19+).

CD4 Low and Normal Blood Controls

The CD4 Normal Blood Control contains CD4+ T cells in the normal range (410-1590 cells/Ál) and the CD4 Low Blood Control contains CD4+ T cells in a low range (200-300 cells/Ál). These controls can be purchased separately or as a pair.

If you would like to be alerted when a new batch of Blood Controls are released, please contact us.

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