The NIR IVFC system used in this study was established by ourselves and equipped with two lasers: a 785-nm laser used to excite NIR fluorescent dyes, and a 488-nm laser used to excite GFP. Once a fluorescence cell in the blood passed through the laser slit, the fluorescence of both channels would be excited and detected at the same time. After the GFP-labeled leukemia cells were transplanted in the mice for 5 days, the mice were injected with IR-26 by tail vein. IVFC for tracking fluorescent cells in peripheral blood of mice was performed according to our previous protocols (24). Briefly, after the mouse was anesthetized with 1% sodium pentobarbital solution, it was placed on the platform, and the GFP and NIR fluorescence information in peripheral blood of mouse was recorded and analyzed using IVFC. The first measurements were acquired within 15 min after the first injection of IR-26. Additional measurements were acquired at the same vessel location.

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