Imaging station, by Kodak - Product details

1

In Vivo NIRF Imaging of Xenograft Tumors

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Example 8

K9TCC-Pu-In and 5637 bladder cancer and SKOV-3 ovarian cancer xenograft mice models with subcutaneous tumor of an approximate 8˜10 mm diameter were subjected to in vivo NIRF optical imaging. At different time points post injection of nano-porphorin micelles, mice were scanned using a Kodak multimodal imaging system IS2000MM with an excitation bandpass filter at 625 nm and an emission at 700 nm. The mice were anaesthetized by intraperitoneal injection of pentobarbital (60 mg/kg) before each imaging. After in vivo imaging, animals were euthanized by CO₂overdose at 24, 48 and 72 h after injection. Tumors and major organs were excised and imaged with the Kodak imaging station.

NIRF imaging studies was performed in the orthotopic xenograft model in NSG mice. Briefly, female NSG mice bearing orthotopic human bladder cancers or normal NSG mice were intravesically injected with 30 μl of PLZ4-NP via urethra under general anesthesia. After 2 hr of incubation, the bladder was isolated for whole body in vivo imaging using Kodak imaging system.

US10238750B2. Porphyrin modified telodendrimers (2019-03-26). The Regents of the University of California [US]. Inventors: Kit Lam [US], Yuanpei Li [US], Chongxian Pan [US].

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In vivo NIRF Optical Imaging of Tumor Xenografts

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Example 8

K9TCC-Pu-In and 5637 bladder cancer and SKOV-3 ovarian cancer xenograft mice models with subcutaneous tumor of an approximate 8-10 mm diameter were subjected to in vivo NIRF optical imaging. At different time points post injection of nano-porphorin micelles, mice were scanned using a Kodak multimodal imaging system IS2000MM with an excitation bandpass filter at 625 nm and an emission at 700 nm. The mice were anaesthetized by intraperitoneal injection of pentobarbital (60 mg/kg) before each imaging. After in vivo imaging, animals were euthanized by CO₂overdose at 24, 48 and 72 h after injection. Tumors and major organs were excised and imaged with the Kodak imaging station.

NIRF imaging studies was performed in the orthotopic xenograft model in NSG mice. Briefly, female NSG mice bearing orthotopic human bladder cancers or normal NSG mice were intravesically injected with 30 μl of PLZ4-NP via urethra under general anesthesia. After 2 hr of incubation, the bladder was isolated for whole body in vivo imaging using Kodak imaging system.

US11219692B2. Porphyrin modified telodendrimers (2022-01-11). THE REGENTS OF THE UNIVERSITY OF CALIFORNIA [US]. Inventors: Kit S. Lam [US], Yuanpei Li [US], Chong-Xian Pan [US].

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In Vivo Biodistribution of HEV-VLP Nanoparticles

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Female SPF BALB/c mice, 6–8 weeks of age, were purchased from Charles River (CA, USA) and kept under pathogen-free conditions according to Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) guidelines and were allowed to acclimatize for at least 4 days prior to any experiments. MDA-MB-231 breast cancer cells (5 × 10⁵) were injected into the mammary fat pad of female nude mice. All animal experiments were performed in compliance with institutional guidelines and according to protocol no. 17222 approved by the Animal Use and Care Administrative Advisory Committee at the University of California, Davis. MDA-MB-231 tumors bearing mice were intravenously injected through the tail vein with 4 nmol/l Cy5.5 fluorescent-labeled HEV-573C-VLPs (VLP-Cy5.5) and LXY30-conjugated HEV-573C-VLPs (LXY30-VLP-Cy5.5), respectively. At different time point (1, 6, 24 and 48 h post-injection), the mice were scanned with Kodak imaging system IS2000MM according to the procedure described previously [17 (link)]. Imaging was collected with an excitation bandpass filter at 625 nm and an emission at 700 nm at exposure time 30 s per image. After in vivo imaging, animals were euthanized by CO₂ overdose at 72 h after injection. Tumors, organs and muscle tissues were excised and imaged with Kodak imaging station.

Chen C.C., Xing L., Stark M., Ou T., Holla P., Xiao K., Kamita S.G., Hammock B.D., Lam K, & Cheng R.H. (2016). Chemically activatable viral capsid functionalized for cancer targeting. Nanomedicine, 11(4), 377-390.

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In Vivo Optical Imaging of Tumor Nanoparticles

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After transgenic mice (10-12 weeks) and nude mice developed established tumors (6-10 mm in diameter), they were subjected to in vivo NIRF imaging by injecting 100 μL of NPs via tail vein. At different time points post-injection of NPs, mice were scanned using a Kodak multimodal imaging system IS2000MM with an excitation bandpass filter at 625/20 nm and an emission at 700/35 nm under anaesthesia. After in vivo imaging, animals were euthanized. Tumors and major organs were excised and imaged with the Kodak imaging station. To monitor the kinetics of biodistribution at the tissue level, SKOV3 tumor-bearing mice were given 100 μL of CNPs via tail vein and euthanized at 1, 24, or 48 hrs. Right before euthanasia, Dextran-FITC solution was injected intravenously to locate blood vessel. Tumors were harvested and fixed in the cold formalin on ice. A fresh cross-section was made for Large-Scale-Imaging (LSI) laser scanning confocal microscope imaging. Additionally, lungs with metastatic lesions were also collected from older transgenic mice (20-24 weeks) and fixed in cold formalin for one hour. The lung surface was imaged by LSI laser scanning confocal microscopy. After imaging, lungs were subjected to histopathological evaluation. In some cases, the blood of the mice were drawn and measured at predetermined time points.

Li Y., Lin T.Y., Luo Y., Liu Q., Xiao W., Guo W., Lac D., Zhang H., Feng C., Wachsmann-Hogiu S., Walton J.H., Cherry S.R., Rowland D.J., Kukis D., Pan C, & Lam K.S. (2014). A Smart and Versatile Theranostic Nanomedicine Platform based on Nanoporphyrin. Nature communications, 5, 4712.

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5

In Vivo Optical Imaging of Tumor Nanoparticles

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After transgenic mice (10-12 weeks) and nude mice developed established tumors (6-10 mm in diameter), they were subjected to in vivo NIRF imaging by injecting 100 μL of NPs via tail vein. At different time points post-injection of NPs, mice were scanned using a Kodak multimodal imaging system IS2000MM with an excitation bandpass filter at 625/20 nm and an emission at 700/35 nm under anaesthesia. After in vivo imaging, animals were euthanized. Tumors and major organs were excised and imaged with the Kodak imaging station. To monitor the kinetics of biodistribution at the tissue level, SKOV3 tumor-bearing mice were given 100 μL of CNPs via tail vein and euthanized at 1, 24, or 48 hrs. Right before euthanasia, Dextran-FITC solution was injected intravenously to locate blood vessel. Tumors were harvested and fixed in the cold formalin on ice. A fresh cross-section was made for Large-Scale-Imaging (LSI) laser scanning confocal microscope imaging. Additionally, lungs with metastatic lesions were also collected from older transgenic mice (20-24 weeks) and fixed in cold formalin for one hour. The lung surface was imaged by LSI laser scanning confocal microscopy. After imaging, lungs were subjected to histopathological evaluation. In some cases, the blood of the mice were drawn and measured at predetermined time points.

Li Y., Lin T.Y., Luo Y., Liu Q., Xiao W., Guo W., Lac D., Zhang H., Feng C., Wachsmann-Hogiu S., Walton J.H., Cherry S.R., Rowland D.J., Kukis D., Pan C, & Lam K.S. (2014). A Smart and Versatile Theranostic Nanomedicine Platform based on Nanoporphyrin. Nature communications, 5, 4712.

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6

HPV Detection in Cell and Tumor Samples

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DNA extraction was done from treated and untreated cell samples as well as mouse tumor samples using Promega DNA extraction Kits (Madison, WI) according to manufacturer's protocol. The polymerase chain reaction (PCR) was used to detect the expression of human papilloma virus (HPV) gene as per the manufacturer's protocol using Promega-Master Mix Kit (Madison, WI). The analyses were performed on 500 ng of purified DNA of treated and untreated cell and tumor samples as described below. PCR reaction contained 500 ng of DNA, 4 μL of Master Mix, and 4 μL of each consensus primer (MY11, MY09, β-actin reverse and β-actin forward). Thirty cycles of start denaturation (95°C for 4 min), denaturation (94°C-1 min), annealing (55°C-1 min), and extension (72°C-1.5 min) were performed on a BioRad thermocycler. β-actin was used as the DNA loading control. After the completion of the reaction, 10 μL each of all PCR products were resolved on 1.5% agarose gel with ethidium bromide against 100 bp DNA ladder. The image was captured under UV illumination using Kodak Imaging Station (Kodak, Rochester, NY). The primer sequence of MY11 is DNA 5′ to 3′-GCM CAG GGW CAT AAY AAT GG and MY09 is DNA 5′ to 3′-CGT CCM ARR GGA WAG TGA TC. The sequence of β-actin forward primer is AAC TGG GAC GAC ATC GAG AA and the sequence of β-actin reverse primer is AGA GGC GTA CAG GGA TAG CA.

Smith J.A., Mathew L., Gaikwad A., Rech B., Burney M.N., Faro J.P., Lucci JA I.I.I., Bai Y., Olsen R.J, & Byrd T.T. (2019). From Bench to Bedside: Evaluation of AHCC Supplementation to Modulate the Host Immunity to Clear High-Risk Human Papillomavirus Infections. Frontiers in Oncology, 9, 173.

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7

Protein Expression Analysis in Transgenic Lettuce

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The expression of the target gene was detected by western blotting. The total protein from control and transgenic lettuce plants that showed a hybridization signal in the Southern blot was extracted using a Plant Protein Extraction Kit (ComWin, China). The protein samples were separated on a 10% Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) gel and electro-transferred onto a Polyvinylidene Fluoride (PVDF) membrane in an electroblotting transfer buffer. The membranes were blocked with 5% non-fat milk powder in Tris Buffered Saline Tween (TBST) with gentle agitation overnight to prevent non-specific antibody reaction. The membrane was incubated at 25°C for 2 h in a 1:800 dilution of Anti-His Tag Monoclonal Antibody (ComWin, China) in a TBST antibody dilution buffer and washed five to six times with TBST buffer. Then, the membrane was incubated for 2 h in a 1:50,000 dilution of goat anti-mouse antibody labeled with horseradish peroxidase (Boster, China) in TBST buffer and washed five to six times using TBST buffer. Antibody binding was detected by incubation with an ECL substrate solution, following the manufacturer's instructions. An image analysis of the western blot was conducted using a Kodak Imaging Station (Kodak, USA).

Song D., Xiong X., Tu W.F., Yao W., Liang H.W., Chen F.J, & He Z.Q. (2017). Transfer and expression of the rabbit defensin NP-1 gene in lettuce (Lactuca sativa). Genetics and molecular research : GMR, 16(1).

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Imaging station

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7 protocols using imaging station

In Vivo NIRF Imaging of Xenograft Tumors

In vivo NIRF Optical Imaging of Tumor Xenografts

In Vivo Biodistribution of HEV-VLP Nanoparticles

In Vivo Optical Imaging of Tumor Nanoparticles

In Vivo Optical Imaging of Tumor Nanoparticles

HPV Detection in Cell and Tumor Samples

Protein Expression Analysis in Transgenic Lettuce

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