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. 2005 Oct-Dec;4(4):415-6.
doi: 10.2310/7290.2005.05148.

Real-time detection of circulating apoptotic cells by in vivo flow cytometry

Xunbin Wei  1 , Dorothy A Sipkins, Costas M Pitsillides, John Novak, Irene Georgakoudi, Charles P Lin
Affiliations

Real-time detection of circulating apoptotic cells by in vivo flow cytometry

Xunbin Wei et al. Mol Imaging. 2005 Oct-Dec.
No abstract available

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Figures

Figure 1
Figure 1
Detection of circulating apoptotic cells with in vivo flow cytometry. (Solid circle) Camptothecin-treated MatLyLu cells (~1 × 106) in 100 μL PBS were labeled with annexin-V probe conjugated to AF647 (Molecular Probes; 2.95 μg), washed, and injected into the mouse tail vein. Mice were anesthetized and placed on a heated stage (37°C). An appropriate vessel (usually a small artery) for obtaining measurements was chosen in the ear. Fluorescence signal was recorded as individual labeled cells passed through a slit of He – Ne laser (633 nm) focused across the blood vessel. Confocal detection of the excited fluorescence enabled continuous monitoring of labeled cells flowing through the vessel over time. Signal was detected by a photomultiplier tube and digitized for analysis with Matlab software developed in-house [ – 3]. The number of annexin-V+ cells in the circulation was measured by taking sequential 60-sec data traces and plotted as a function of time after injection. Inset shows a representative data trace where two annexin-V+ cells could be detected during the 1-sec time period (A.U. denotes arbitrary unit). (Open square) Number of annexin-V+ cells detected in the mouse circulation when unlabeled, camptothecin-treated MatLyLu cells (~1 × 106) in 100 μL PBS were injected, followed immediately by injection of the annexin-V probe (11.8 μg). (Cross) Control experiments with an injection of the annexin-V probe alone.
Figure 2
Figure 2
(Solid circles) Time course of annexin-V+ cell count following injection of ~1 million untreated MatLyLu cells. Injection of the annexin-V probe at each time point (10 min, 1 hr, 2 hr, 3 hr, 5.5 hr, and 8.5 hr) was necessary because of its short circulation time [8]. Measurement was taken 5 min after each probe injection to allow binding of the probe to the apoptotic cells. (Standard errors were shown; n = 3). The experiments using ~1 million LNCaP prostate cancer cells and ~3 million leukemic Nalm-6 cells gave similar results (open squares and solid triangles, respectively; standard errors were not shown for the clarity of the figure).
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References

    1. Novak J, Georgakoudi I, Wei X, Prossin A, Lin CP. In vivo flow cytometer for real-time detection and quantification of circulating cells. Opt Lett. 2004;29:77–79. - PMC - PubMed
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