Ivis lumina k series 3

ASCs were cultured for several days until they reached confluence and then were replaced with 5 mL of DLU2-NPs diluted 640-, 320-, or 160-fold with medium. After incubation for 4 h, ASCs were washed twice with medium and collected in 1.5 mL centrifuge tubes. The fluorescence images of ASCs with DLU2-NPs in 1.5 mL centrifuge tubes were taken using the IVIS Lumina K Series III (PerkinElmer Inc, Waltham, MA, USA; excitation filter: 460–620 nm, emission filter: 520, 570, and 710 nm long-pass). In vitro MR images of ASCs with DLU2-NPs in 1.5 mL centrifuge tubes were taken using the MR VivoLVA (DS Pharma Biomedical Co., Ltd., Osaka, Japan). Regarding the spin echo, the images were obtained with repetition time (TR) = 500.0 ms and echo time (TE) = 9.0 ms, or TR = 2000.0 ms and TE = 69.0 ms and field of view = 60 × 60 mm.

All mice were imaged on an IVIS Lumina K Series III living animal biophoton imaging system (PerkinElmer) immediately after infection. Signals were collected from the defined region of interest (ROI) using the contour ROI tool, and the average flux intensity (p/s/cm²/sr) was analysed using Living Image Software 4.7.3. Each group of mice was gently anaesthetised with isoflurane through a small animal anaesthesia machine (RWD‐R540). At 24 h after infection, the infected mice in each group were treated according to the dosage of antibiotics or (and) LFU for 3 days, and bioluminescence imaging was performed on all mice every day to quantify the bacterial load.

Athymic nude mice (BALB/c, 4–6-week-old) and C57BL/6 mice (5–6-week-old) were obtained from Model Organisms (Shanghai, China). Renal cancer cells, including ACHN and RENCA treated with DMSO, ICI, or cuprotosis inducer reagents (Elesclomol + Cucl2), were injected into right flank of mice (1.0 × 10⁶ cells per mice), respectively. BALB/c mice inoculated with RENCA were observed after 12 days; then those mice were randomized divided into three groups receiving DMSO, cuprotosis inducer reagents at concentration of 2 and 5 μmol/L, and tumor was identified at 24 days by bioluminescent imaging by IVIS Lumina K series III (PerkinElmer Inc, Hopkinton, MA, USA). After the xenografts reached a size of approximately 45 mm³, C57BL/6 mice were randomized into four groups receiving DMSO, anti-PD-1mAB, cuprotosis inducer reagents, and combined therapy (anti-PD-1 + cuprotosis inducer reagents) by intraperitoneal (i.p., given at days 14, 17, 20), respectively. After one week, all mice treated with those agents were euthanized and peripheral blood were harvested to quantity the percentage of CD45 + CD8 + T cells among different groups by flow cytometry. All those procedures were approved by the Institutional Animal Care and Use Committee at Navel Medical University.

Six male A129 mice (7–8 weeks old) were randomly divided into two groups. Mice in each group (n = 3) were injected intraperitoneally with Z2-Cy5 (100 μg in 100 μl PBS) or PBS (vehicle in 100 μl PBS) as control. After anesthetization with pentobarbital sodium, mice were imaged by the IVIS Lumina K series III in vivo imaging system from PerkinElmer (Waltham, MA, United States) for 1 h. To determine the distribution of Z2-Cy5 in the testicular tissue, mice were sacrificed using pentobarbital sodium, and all testes and epididymides were removed for imaging. The radiant efficiency (ps^–1cm^–2sr^–1) (μW^–1cm^–2) in mouse body and testis and epididymis was calculated by Living Image 4.5.5 software.

R8-ZZC (ZZC:250 nM, R8:250 μM) in PBS or transduction medium (DMEM/F12 containing 2% FBS and 100 U/mL penicillin/streptomycin) was incubated for various length of time (0, 1, 3, 5 and 7 days) at 37 °C. The phase and fluorescence images were observed by 8 mega pixel digital camera and the intensities were measured using an IVIS Lumina K Series III (PerkinElmer, Inc., Massachusetts, USA).