Stability of canine and feline cerebrospinal fluid samples regarding total cell count and cell populations stored in "TransFix®/EDTA CSF sample storage tubes"
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Laura Meier1, Regina Carlson2, Jasmin Neßler2
1Department of Small Animal Medicine and Surgery, University of Veterinary Medicine, Hannover, Germany. laura.meier@tiho-hannover.de.
Insights
Canine and feline cerebrospinal fluid (CSF) samples stored in TransFix® tubes allow for extended flow cytometry analysis of lymphocytes and granulocytes up to three days. However, standard cell counts and morphology assessments require fresh samples.
Area of Science:
- Veterinary Medicine
- Clinical Pathology
- Laboratory Diagnostics
Background:
- Cerebrospinal fluid (CSF) leukocyte degeneration necessitates prompt laboratory examination post-collection.
- Rapid degeneration limits routine and flow cytometry analysis of CSF samples.
Purpose of the Study:
- To evaluate the efficacy of "TransFix®/EDTA CSF Sample Storage Tubes" for preserving canine and feline CSF samples.
- To determine the feasibility of routine and flow cytometry examinations up to 3 days post-sampling.
Main Methods:
- Storage of canine and feline CSF samples in "TransFix®/EDTA CSF Sample Storage Tubes".
- Assessment of leukocyte viability and morphology using Türk's solution and cytospin preparations.
- Flow cytometry analysis using antibodies against various cell surface markers (CD3, CD4, CD21, CD11b, CD45, CD14).
Main Results:
- Leukocyte staining and morphology assessment were compromised in stored samples.
- Flow cytometry showed significantly higher cell counts in TransFix®-stored samples compared to untreated samples.
- Antibody binding for CD3, CD4, CD21, CD11b, and CD45 was effective, enabling cell population differentiation, but CD14+ monocytes were undetectable.
Conclusions:
- "TransFix®/EDTA CSF Sample Storage Tubes" are suitable for extended storage of CSF for flow cytometry of lymphocytes and granulocytes.
- Monocyte detection in CSF is not feasible after storage in TransFix® tubes.
- Fresh CSF samples are essential for standard microscopic cell counting and morphological evaluation.
Background:
Because of fast leucocyte degeneration in cerebrospinal fluid (CSF) laboratory examinations of CSF samples should be performed approximately within 30 min after withdrawal. This study examines the storage of canine and feline CSF samples in "TransFix®/EDTA CSF Sample Storage Tubes" (Cytomark, Buckingham, UK) for preventing leucocytes from degeneration, so that routine and flow cytometry examinations are feasible up to 3 days after sampling.
Results:
After storage in TransFix® tubes, leukocytes could not be adequately stained with Türk's solution and differentiating between erythrocytes and leukocytes was cumbersome. In addition, the cell morphology could not be sufficiently assessed on cytospin preparations because of shrunken leukocytes and indistinct cell nuclei. In contrast, by flow cytometry, a significantly higher cell count was measured over the entire study period in the samples stored in TransFix® tubes compared to the untreated samples. The antibodies (AB) against CD3, CD4 and CD21, against CD11b and against CD45 showed a good binding strength and thus enabled a good differentiation of cell populations. However, after storage in the TransFix® tubes, monocytes were no longer detectable using an AB against CD14.
Conclusion:
Based on these results, "TransFix®/EDTA CSF Sample Storage Tubes" can be used for extended storage prior to flow cytometric analysis of lymphocytes and granulocytes in CSF samples but not for detecting monocytes. However, standard examinations, such as microscopic cell counting and morphological cell assessment should be performed on fresh CSF samples.
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