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A method for tracking the migration of blood lymphocytes
W N Andrade1, M G Johnston, J B Hay
1Department of Pathology, University of Toronto, Ontario, Canada.
Immunological Investigations
|September 1, 1996
Summary
Researchers developed a new method to label whole blood with fluorescein isothiocyanate (FITC) for tracking sheep lymphocytes. This technique allows for efficient labeling of large lymphocyte numbers for migration studies.
Area of Science:
- Immunology
- Cell Biology
- Animal Science
Background:
- Studying lymphocyte migration is crucial for understanding immune responses.
- Previous methods for labeling lymphocytes often result in significant cell loss and require complex purification steps.
- Accurate quantification of lymphocyte recirculation necessitates labeling a large number of cells (10^8-10^9).
Purpose of the Study:
- To develop an efficient method for labeling lymphocytes in whole blood for migration studies in sheep.
- To overcome limitations of cell loss and in vitro manipulation associated with traditional lymphocyte purification techniques.
- To enable the tracking of a sufficient number of lymphocytes for reliable detection and quantification in blood and lymph.
Main Methods:
- Whole blood samples from sheep were labeled directly with fluorescein isothiocyanate (FITC).
- Labeled cellular components, including lymphocytes, were reinjected intravenously into the animals.
- Lymphocytes were distinguished from other labeled cells using flow cytometry.
Main Results:
- The FITC labeling method successfully labeled between 2.4-12.4 x 10^8 lymphocytes.
- Labeled lymphocytes were detected in blood and lymph for at least 10 days post-injection.
- Recovery of FITC-labeled lymphocytes in efferent lymph was comparable to other labeling methods.
Conclusions:
- Labeling whole blood with FITC is an effective and efficient method for tracking lymphocyte migration in sheep.
- This technique minimizes cell loss and in vitro manipulation, ensuring lymphocyte viability and recirculation.
- The method is suitable for studies involving cannulated lymphatic vessels and flow cytometry analysis of lymphoid organs, but not for histological localization studies.