Figure 1
Figure 1. In vivo monitoring of hematopoietic reconstitution. (A) Schematic drawing represents the fluorescence confocal endoscope system (CellVizio, MKT) equipped with the S300 flexible microprobe containing 10 000 optical fibers that carry light from a continuous laser source at 488 nm to the living tissue. The distribution of transplanted fluorescent hematopoietic cells was monitored through the femur of living mice from the knee to the femoral head using video data acquisition software (IC-Viewer, MKT). (B) At 24 hours after irradiation at 10 Gy, C57BL/6 recipient mice were transplanted with 1000 GFP+ LSKCD34− sorted cells. Nine (top panel) and 14 (bottom panel) days after transplantation, the GFP+ hematopoietic cell tracking was performed. Pictures representative of GFP+ hematopoietic cells present in the knee, diaphysis, and femoral head areas are shown. These pictures represented frames of 30 to 50 seconds acquired videos over the length of the femur. Scale bar represents 50 μm. The graphics shown below the pictures represent the mean of the relative fluorescence units detected over the time in the femur by the 10 000 optical fibers per individual video frame. The vertical strokes indicate different positions of the microprobe tip in the femoral cavity. Only relative fluorescence units in excess of a value of 60 is considered a positive signal. Note the scale difference between the relative fluorescence units detected 9 and 14 days after transplantation. These data are representative of 5 to 10 independent experiments. (C) Quantification and fluorescence mean of the images obtained 9 and 14 days after transplantation of 1000 GFP+ LSKCD34− hematopoietic cells. Seventy-five, 200, and 100 video frames were studied in the knee, the diaphysis, and the femoral head, respectively. The number of GFP+ frames and the fluorescence mean of each frame are indicated for each femoral compartment. These data are representative of 5 to 10 independent experiments. Statistical analysis was performed with the 1-way analysis of variance test.

In vivo monitoring of hematopoietic reconstitution. (A) Schematic drawing represents the fluorescence confocal endoscope system (CellVizio, MKT) equipped with the S300 flexible microprobe containing 10 000 optical fibers that carry light from a continuous laser source at 488 nm to the living tissue. The distribution of transplanted fluorescent hematopoietic cells was monitored through the femur of living mice from the knee to the femoral head using video data acquisition software (IC-Viewer, MKT). (B) At 24 hours after irradiation at 10 Gy, C57BL/6 recipient mice were transplanted with 1000 GFP+ LSKCD34− sorted cells. Nine (top panel) and 14 (bottom panel) days after transplantation, the GFP+ hematopoietic cell tracking was performed. Pictures representative of GFP+ hematopoietic cells present in the knee, diaphysis, and femoral head areas are shown. These pictures represented frames of 30 to 50 seconds acquired videos over the length of the femur. Scale bar represents 50 μm. The graphics shown below the pictures represent the mean of the relative fluorescence units detected over the time in the femur by the 10 000 optical fibers per individual video frame. The vertical strokes indicate different positions of the microprobe tip in the femoral cavity. Only relative fluorescence units in excess of a value of 60 is considered a positive signal. Note the scale difference between the relative fluorescence units detected 9 and 14 days after transplantation. These data are representative of 5 to 10 independent experiments. (C) Quantification and fluorescence mean of the images obtained 9 and 14 days after transplantation of 1000 GFP+ LSKCD34− hematopoietic cells. Seventy-five, 200, and 100 video frames were studied in the knee, the diaphysis, and the femoral head, respectively. The number of GFP+ frames and the fluorescence mean of each frame are indicated for each femoral compartment. These data are representative of 5 to 10 independent experiments. Statistical analysis was performed with the 1-way analysis of variance test.

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