Ms fx pro

For the fluorescence imaging of a breast-tumour, an aqueous solution (200 μL) of ICG-labelled Kadcyla (1 mg mL⁻¹) was intravenously injected via a tail vein of the mouse. NIR fluorescence images of the tumour were taken 0, 1, 3, and 5 days after the injection of ICG labelled Kadcyla. Five days after of the probe, ex vivo images of a breast tumour and organs were taken.
For the fluorescence imaging of tumour-apoptosis, Kadcyla (200 μL, 1 mg mL⁻¹) was intravenously injected via a tail vein of breast tumour-bearing mice. Three days after the injection of Kadcyla, ICG–EGFP (or Plum)–Annexin V or ICG–Annexin V (200 μL, 1 mg mL⁻¹) was injected to the mice. Three days after the probes, fluorescence images of a mouse treated were taken.
Fluorescence images were taken using an in vivo fluorescence imaging system (Bruker, MS FX PRO). NIR fluorescence of ICG was observed at 830 ± 20 nm by excitation at 760 nm. Exposure time of the excitation light was 30 s. VIS fluorescence of EGFP and mPlum was observed at 515 ± 20 nm (ex: 470 nm) and 670 ± 20 nm (ex: 590 nm), respectively. Exposure time of the excitation light was 1 s for EGFP emission and 10 s for mPlum emission. Excitation light (400 W Xenon lamp) power was 30 μW cm⁻² at a ventral side of a mouse.

Adult zebrafish (n = 10) was individually exposed immersed for 5 minutes in a becker of 500 mL containing NPMA in concentration 0.050 or 0.025 mg.mL-1, in accordance with protocols described by Charlie-Silva et al. [31 ]. After the time of immersion, the zebrafish were transferred to a becker with 500 mL of clean water to be measured the interaction of NPMA and fish surface. Immediately after anesthesia were analyzed in vivo fluorescence images produced by nanocapsules that were absorbed by zebrafish tissues using a Bruker Xtreme In-Vivo Imaging System (Bruker, Billerica, MA, USA). Fish liver, gill and intestine images confocal intravital microscopy was performed as previously described by Marques et al. [32 ]. The counts were performed on the images captured in the confocal microscope at MS FXPRO (Bruker).

The rostrocaudal macroscopic distribution of AFO-647 along the neuraxis was captured with white-light and a single fluorescent channel (excitation wavelength 630 nm and emission wavelength 700 nm, exposure time of 4 s) using the small animal optical imaging system MS FX PRO (Bruker UK Ltd.). Images were taken from both the dorsal and ventral directions.
Spinal cord axial sections from C2–T4 were imaged with a Zeiss Axio Imager fluorescence microscope (Carl Zeiss Microimaging GmbH, Germany). Immunohistochemistry was used to image the spinal vasculature and identify vessel types. Arterioles were identified as vessels positive for RECA-1 and SMA, whereas venules and capillaries were labelled by RECA-1 only. Blood vessels that had a luminal diameter < 6.5 μm were classified as capillaries. Confocal microscopy (LSM 880, Carl Zeiss Microimaging GmbH, Germany) was used to further characterise vascular structures and the central canal.

For each rat belonging to the five experimental groups, radiographic analyses were performed, using an X-ray machine (Bruker MS FX Pro, Billerica, MA, USA). The X-ray tube was operated at 30 kW, with a current of 6 mA, for 0.01 s, and the source-to-sensor distance was 50 cm. At the end of the experiment, through the radiographs, we estimated the dental alveolar bone level expressed as the distance from the cement–enamel junction (CEJ) to the maximum coronal level of the alveolar bone crest (CEJ bone distance), using IMAGE J processing software (Image J software, National Institutes of Health, Bethesda, MD, USA).

After post fixation, macroscopic white-light and single channel fluorescent images were captured with the in-vivo MS FX PRO (Bruker, Billerica, MA). The fluorescence camera was set at excitation and emission wavelengths of 630 and 700 nm respectively, with an exposure time of 4 s.
Spinal cord axial sections from C2 to T4 were imaged with a Zeiss Axio Imager Z1 fluorescence microscope (Carl Zeiss Microimaging GmbH, Germany) for qualitative and quantitative analysis. The fluorescent microspheres, which have a diameter of 1 μm, were used to verify the location of the injection site as their size prevents significant displacement. SMA- and RECA-1-positive vessels were identified as arterioles. SMA-negative, RECA-1-positive vessels were designated venules or capillaries. Those with largest diameter ≥ 6.5 µm were considered venules, and those < 6.5 µm capillaries. Further delineation of vascular and anatomical structures was undertaken with confocal microscopy (LSM 880, Carl Zeiss Microimaging GmbH, Germany).

Tumor development was monitored using an in vivo imaging device, Bruker model MSFXPRO. Throughout image acquisition, animals were placed in dorsal recumbency and remained anesthetized with inhaled 2% isoflurane in oxygen at 2 L/min. Initially, the skull images were acquired by X-ray. The fluorescence of the labeled cells was evaluated using the excitation (540 nm) and emission (585 nm) of MION-Rh and excitation (405–665 nm) and emission (705 nm) of Qdots (705 nm). The images were acquired and evaluated using multiplex location software.

N-Hydroxysuccinimidyl ester derivatives of organic dyes, Alexa488–NHS ester and ICG–NHS ester were purchased from Thermo Fisher Scientific and Goryo Chemicals (Japan), respectively. Heceptin, Cyramza, and Kadcyla were purchased from Chugai Pharmaceutical Co., Ltd (Tokyo, Japan). Erbitux was purchased from Merk Serono. Anti-human PD-L1 antibody was purchased from Bio Cell. Normal human IgG was purchased from FUJIFILM Wako Pure Chemical Corp. (Japan). Glutathione-coated PbS QDs (E_m: 1250 nm) were prepared by the literature method.^{36 (link)} Intralipid emulsion (20%) was purchased from Sigma-Aldrich. Breast tumour cells (KPL-4) were kindly provided by Dr Kurebayashi (Kawasaki Medical School). Skin tumour cells (A431) were purchased from RIKEN cell bank. Nude mice (five-week-old female BALB/c nu/nu mice) were purchased from SLC Inc (Japan).
Absorption spectra were recorded with a spectrophotometer (Jasco, V-670). Fluorescence spectra were recorded with a spectrofluorometer (NanoLog, HORIBA, Japan). Fluorescence imaging of cancer cells were performed with a fluorescence microscope (BZ-X700, Keyence, Japan). In vivo NIR fluorescence imaging was performed with a fluorescence system (Bruker MS-FX PRO). In vivo SWIR fluorescence imaging was performed with a home-built wide-field microscope system.^4a