Xenogen ivis 200 spectrum camera

MOLM14-Luc cells or CFSE (Invitrogen) labeled primary AML samples were used for cytotoxicity assay as previously described.^{28 (link)} In brief, targets were incubated at the indicated ratios with effector T cells for 4 or 16 h. Killing was calculated either by bioluminescence imaging on a Xenogen IVIS-200 Spectrum camera (PerkinElmer, Hopkinton, MA, USA) or by flow cytometry. For the latter, cells were harvested; Countbright beads and 7-AAD (Invitrogen) were added prior to analysis. Residual live target cells were CFSE+ 7-AAD–. For MDS, T cells were incubated with CD34-selected bone marrow at 1:1 ratio for 4 or 24 h as indicated, and cytotoxicity was then measured by flow cytometry or by fluorescence in situ hybridization using a probe to detect 5q deletion or monosomy 5 (EGR1 (5q31)/ D5S23, D5S721 (5p15.2)).

Luciferase⁺ Nalm6, OPM-2, and K562 cells were incubated at the indicated ratios with effector T cells for 24 hours. The killing was calculated by bioluminescence imaging (BLI) on a Xenogen IVIS-200 Spectrum camera (cat# 124262, PerkinElmer, Hopkinton, MA, USA) as a measure of residual live cells. Ten minutes prior to imaging, samples were treated with 1 µL D-luciferin (30 µg/mL, Gold Biotechnology, St. Louis, MO, USA) per 100 µL sample volume.

Subcutaneous Nalm6 xenografts were established through the subcutaneous injection of 1 x 10⁶ luciferase⁺ Nalm6 cells in PBS. Tumor burden was assessed by BLI using a Xenogen IVIS-200 Spectrum camera (PerkinElmer, Waltham, MA, USA) to confirm engraftment weekly after Nalm6 inoculation. Once the tumor volume reached 50 mm in diameter, mice were then randomized into groups receiving 1) UTD or 2) NIS⁺CART-cells (5 x10⁶ intravenously). Weekly Imaging was performed 10 minutes after the intraperitoneal (IP) injection of 10 µL/g D-luciferin (15 mg/mL, Gold Biotechnology, St.Louis, MO, USA). Bioluminescent images were analyzed using Living Image version 4.4 (PerkinElmer, Waltham, MA, USA). One week after T-cell injection, mice were imaged both with BLI and ¹⁸F-TFB-PET/CT as described below. Mice were euthanized after the PET imaging and the experiment was concluded.