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Ex Vivo Immunophenotypization

Leukocyte activation and function

Differential cell count and determination of eosinophils and neutrophils within bronchoalveolar lavage fluid (BALF), peritoneal lavage (PL) or whole blood leukocyte population are commonly used methods for measurement of inflammatory response in animal models. Manual cell count is time-consuming and prone to errors due to low number of cells and subjectivity in differentiation.

Fidelta has developed a novel flow cytometric method for the detection of murine granulocytes. The method is applicable in every in vivo inflammatory model where identification and count of eosinophils and/or neutrophils in BALF, peritoneal lavage fluid and blood samples is desirable.

Material

  • BALF
  • Peritoneal lavage fluid
  • Whole blood


Available read-outs

  • Immunophenotypization by flow cytometry
  • Absolute cell count
  • Differential cell count

BALF cells immunophenotypization - mouse model of chronic lung eosinophilia

A simple 3-color flow cytometry method using FITC, anti-mouse CD45 and anti-mouse Gr-1 antibody provides the possibility for  determination of eosinophil (white arrow), neutrophil, monocyte/macrophages (yelow arrow) and lymphocyte  subpopulations in BALF, peritoneal lavage fluid and blood. Presented are BALF analysis examples.



BALF subpopulations
Region/Gate Colour
Cell type
FITC
CD45
Gr-1
Blue
Eosinophils
+
bright +
From dim+ to bright +
Magenta
Neutrophils
-
bright +
High +
Green
Monocytes/Macrophages
-
bright +
Partially dim+
Red
Lymphocytes
-
bright +
-

 

BALF cells immunophenotypization - mouse model of acute lung neutrophilia

Simple 3-color flow cytometry method using anti-mosue F4/80, anti-mouse CD45 and anti-mouse Gr-1 antibody provides a possibility for  determination and enumeration of neutrophil, monocyte/macrophage and lymphocyte  subpopulations in BALF. Presented are BALF analysis examples.

 

 

 

BALF subpopulations
Region/Gate Colour
Cell type
CD45
Gr-1
F4/80
Magenta
Neutrophils
bright +
High +
-
Green
Monocytes/Macrophages
bright +
Partially dim+
+
Red
Lymphocytes / Lymphocyte-like leukocytes
bright +
-
+

 

References

Bedner et al. 1999.  Cytometry 36, 77.
Efthimidiais et al. 1996. Cytometry 26, 75.
Leong et al. 2001, J  Am  Chem  Soc. 120,12237.
Polancec et al. 2006. ISAC XIII Congress. 160631-p369
Polancec et al. 2006. ISAC XIII Congress. 160685-p294

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