Caliper ivis

Guinea pigs were killed to sample the acoustic bullae (including the RWM). The acoustic bullae were placed in solution of FBSA and BSA NPs (the preparation of which was described in another paper by the same authors being reviewed) and shaken for 30 minutes at 37°C. The air-dried specimens were observed by SEM.
The penetration of RhB released from the particles was evaluated by living images and microscopes. The same amount of RhB was incubated with FBSA-H particles and BSA NPs in the preparation of FBSA-RhB and BSA-NP-RhB. Guinea pigs were anesthetized, and the RWMs were exposed. The FBSA-RhB and BSA-NP-RhB particles dispersed in phosphate-buffered saline were injected slowly into the bulla of the right and left ear, respectively. The left ear injected with BSA-NP-RhB was the control. An in vivo imaging system (Caliper IVIS^®; PerkinElmer, Waltham, MA, USA) was used to trace the particles at time points of 0 and 48 hours. RWMs were then imaged by fluorescence microscopy to observe the distribution of RhB on them. Several days later, RWMs with particles were observed by SEM.

The in vivo distribution of PPD NPs and PP NPs were evaluated by the 4T1 tumor-bearing Balb/c mice. When the tumors were up to 300 mm³, the mice were randomly divided into two groups. Cy7-labeled PP NPs and PPD NPs were intravenously administrated at an equivalent dose of 2 mg/kg Cy7. Then, the mice were sacrificed at 4 h and 24 h, and the major organs and tumors were imaged under caliper IVIS (PerkinElmer, USA). As such, the tumors were stained with LDLr primary antibody overnight at 4 °C and Alexa Fluor 488 conjugated anti-rabbit IgG secondary antibody, DAPI and then visualized with CLSM [30] (link).

The biodistribution was investigated in B16-F10 melanoma bearing mice. When the tumor volume reached about 500 mm³, DiR solution and DiR labeled liposomes were administered intravenously at a dose equivalent of 2 mg/kg. After administration for 4, 12 and 24 h, mice were sacrificed. The major organs and tumors were collected for ex vivo imaging by Caliper IVIS (Perkin Elmer, USA).