In vivo fluorescence imaging

A． Preparation of GF-Ala-Cy5.5

1.      Add GF-Ala (0.5 mmol/L, 5 ml), EDC, and NHS (molar ratio, 1:1:2.5) in flask.

2.      Stir at room temperature for 30 minutes (nitrogen protection).

3.      Add 2 mg Sulfo-Cyanine5.5 amine in flask.

4.      Stir at room temperature for 4 hours (nitrogen protection).

5.      Dialyze for several days (dialysis bag, 3500 D).

6.      The concentration of GF-Ala-Cy5.5, determined as Gd element concentration, is measured by ICP-MS.

B． Fluorescence imaging

1.      Mice are injected with GF-Ala–Cy5.5 (1.2 mM, 5 ml/kg, intraperitoneal injection).

2.      Mice are anesthetized, sacrificed, and harvested with heart, liver, spleen, lung, kidney, pancreas, and intestine on the 1st, 5th, 12th, and 30th day.

3.      Using fluorescent imaging system (Spectrum CT, PerkinElmer, USA) to image (excitation wavelength: 675 nm; emission wavelength: 720 nm)

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