Contributions
Abstract: PB1859
Type: Publication Only
Background
In the fluorescence lifetime imaging (FLIM) technique, the image contrast is created by determining the delay of the fluorescence photon emission at each pixel of the image and transforming it in pseudo-colors. This delay, also called lifetime depends on the type of molecules and their physicochemical characteristics.
Aims
We investigated the utility of this technique for the characterization of erythropoietic cell line and changes in the solubility of hemoglobin.
Methods
we used unstained BM smears of 24 normal BM and 8 megaloblastic anemia patients and unstained peripheral blood smears of 10 patients with sickle cell anemia. Images were captured by a confocal microscope with a HPM-100-40-Hybrid detector and excitation at 405 nm (diode laser,80 MHz). In order to create equivalent images of the cytological smears, pseudo-colors were attributed to different lifetime ranges. Images were compared with May-Grünwald-Giemsa (MGG) stained smears.
Results
FLIM created highly contrasted images, where different cell types could be easily recognized by their similarity with MGG images. Erythrocytes exhibited the shortest lifetimes (210.4 ± 42.1 ps). Normal shaped erythrocytes in smears of sickle cell patients showed similar values (214.6 ± 3.1 ps), whereas crenated erythrocytes as well as drepanocytes revealed significantly elevated values (314.2 ± 66.7 ps and 312.5 ± 67.0 ps respectively).
Conclusion
The FLIM technique is easily applicable on unstained routine smears and revealed images of good quality permitting cell identification. It allowed also to distinguish between erythroid and myeloid precursors cells and indicates the major physico-chemical changes during the process of falcization.
Session topic: 23. Hematopoiesis, stem cells and microenvironment
Keyword(s): Erythroid lineage, Erythrocyte, Bone Marrow, Granulopoiesis
Abstract: PB1859
Type: Publication Only
Background
In the fluorescence lifetime imaging (FLIM) technique, the image contrast is created by determining the delay of the fluorescence photon emission at each pixel of the image and transforming it in pseudo-colors. This delay, also called lifetime depends on the type of molecules and their physicochemical characteristics.
Aims
We investigated the utility of this technique for the characterization of erythropoietic cell line and changes in the solubility of hemoglobin.
Methods
we used unstained BM smears of 24 normal BM and 8 megaloblastic anemia patients and unstained peripheral blood smears of 10 patients with sickle cell anemia. Images were captured by a confocal microscope with a HPM-100-40-Hybrid detector and excitation at 405 nm (diode laser,80 MHz). In order to create equivalent images of the cytological smears, pseudo-colors were attributed to different lifetime ranges. Images were compared with May-Grünwald-Giemsa (MGG) stained smears.
Results
FLIM created highly contrasted images, where different cell types could be easily recognized by their similarity with MGG images. Erythrocytes exhibited the shortest lifetimes (210.4 ± 42.1 ps). Normal shaped erythrocytes in smears of sickle cell patients showed similar values (214.6 ± 3.1 ps), whereas crenated erythrocytes as well as drepanocytes revealed significantly elevated values (314.2 ± 66.7 ps and 312.5 ± 67.0 ps respectively).
Conclusion
The FLIM technique is easily applicable on unstained routine smears and revealed images of good quality permitting cell identification. It allowed also to distinguish between erythroid and myeloid precursors cells and indicates the major physico-chemical changes during the process of falcization.
Session topic: 23. Hematopoiesis, stem cells and microenvironment
Keyword(s): Erythroid lineage, Erythrocyte, Bone Marrow, Granulopoiesis