Applications of TransFix: Stabilisation of Bronchoalveolar lavage fluid (BALF)
Independent research involving the use of TransFix to stabilise BALF samples has been published, including the use of TransFix for the enumeration of rat leucocytes present in BALF (case study 1).
Benefits of TransFix for Bronchoalveolar lavage fluid
As with many paucicellular samples, BAL fluid (BALF) can be challenging to analyse because of the low cell numbers present. By stabilising with TransFix, cell degradation is reduced preventing any further loss of cell number.
Please note that Cytomark have not validated the use of TransFix for BALF for IVD use.
Case Study 1: Effects of inhaled nano-TiO2 aerosols showing two distinct agglomeration states on rat lungs
The study examined inflammation resulting from the inhalation of nano-Ti02 aerosols by rats. After exposure to inhalation the rats were anaesthetized with isoflurane and sacrificed by exsanguination. Fluids from the BAL (BALF) were collected with 0.9% saline. The lungs were washed with a total of 21 ml of 0.9 % PBS in 5 steps. These samples were centrifuged to isolate the cells and the cells from the five washes combined. The cell pellet was resuspended in 1 ml of 0.9% PBS containing 1% BSA and 200ul of TransFix. TransFix stabilized cell fractions were examined using a range of techniques to determine the degree of inflammation resulting from inhalation of nano-Ti2 aerosols. The techniques used to examine the TransFix stabilized leucocytes included; total cell counts using a haemocytometer, differential cell counts for lymphocytes using the cytospin cell staining method and inflammation markers were quantified (IL-1 , IL-6, TNF-, MIP-1 and MCP-1) using the Bio-Plex Luminex assay.
Reference: A. Noel, K. Maghni, C. Dion, K.J. Wilkinson, S. Halle, R. Tardif, G. Truchon (2012) Effects of nano-TiO 2 aerosols showing two distinct agglomeration states on rat lungs. Toxicology Letters 214(2): 109-119. https://doi.org/10.1016/j.toxlet.2012.08.019
Case Study 2: Study of body fluid samples using flow cytometry: Six years of experience at the Hospital Universitario San Ignacio – Pontificia Universidad Javeriana, Bogota – Colombia
The Authors discuss findings from 6 years of experience in implementing flow cytometry in Columbia. The authors used TransFix to stabilise all sample, consisting of mainly cerebrospinal fluid, but also bronchoalveolar lavage, pleural fluid, pericardial fluid and ascite fluid from patients with acute and chronic leukaemia, myelodysplastic syndromes, lymphomas, myeloma, autoimmune diseases, immunodeficiencies and solid tumours. The Authors final conclude that this work represents the first report at the national level supporting TransFix implementation in pre-analytical FCM studies of all body fluids that are processed in the clinical practice.
Reference: A. Campos, L. Trujillo, D. López, L. Beltrán , E. Arias, G. Vélez, A. Infante, et al. Study of body fluid samples using flow cytometry: Six years of experience at the Hospital Universitario San Ignacio – Pontificia Universidad Javeriana, Bogota – Colombia. Universitas Scientiarum, 22(2): 123 – 143, 2017. doi: 10.11144/Javeriana.SC22-2.sobf
TransFix is available in 1ml or 20ml vials, or in TransFix Sample Storage Tubes which contain 0.2ml TransFix for the addition of 1ml of sample. If you have any questions regarding the use of TransFix for Bronchoalveolar lavage fluid (BALF), please contact us.