The Aβ-injection AD model was established according to the published methods [80 (link)]. Aβ42 (1 mg/ml) was pre-incubated in artificial cerebrospinal fluid at 37 °C for 7 days. Mice were anesthetized and fixed on a brain stereotactic fixation device for surgery. Pre-incubated Aβ42 (5 μl) was slowly injected into the right hippocampal region. After 2 weeks of recovery, AD mice were randomly grouped and sham-operated mice were used as healthy control. One hundred microliters of Cy5-labeled NPs (PDA and PDA@K, 30 mg/kg) were administrated intravenously at the same concentration of Cy5. At 2, 4, 8, 12, and 24 h post-administration, mice were anesthetized and imaged using the Lumina III Imaging System (PerkinElmer, USA). At 24 h, the mice were sacrificed, and their major organs and brains were separated for ex vivo fluorescence imaging. After that, the organs and brains were fixed with 4% paraformaldehyde, dehydrated in sucrose, embedded by tissue-tek O.C.T compound (Sakura Finetek, USA), and then sectioned (Leica CM1950, Germany). All the slices were stained with DAPI (1 μg/ml) and observed under a confocal microscope.
Lumina 3 imaging system
Manufactured by PerkinElmer
Sourced in United States
The Lumina III Imaging System is a versatile lab equipment designed for imaging and quantification of various biological samples. It provides high-resolution, sensitive detection capabilities for a range of applications.
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13 protocols using lumina 3 imaging system
1
Aβ-Induced Alzheimer's Disease Model Protocol
2
Tumor-targeted Fluorescence Imaging
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100 μL of 4T1 cells (2 × 105) were subcutaneously injected to the left mammary fat pads of female BALB/c nude mouse. 7 days later, mice with tumor around 100 mm3 were randomly divided into three groups (n = 3). Two groups of mice were given d-SN38@NPs and d-SN38@NPs/iRGD, respectively. The last group of mice was intravenously administrated with d-SN38@NPs/iRGD, while the tumor sites of the mice were irradiated with 650 nm laser at 1 h before every imaging point. The fluorescence signal (Ex 640 nm, Em 710 nm) was determined at 4 and 24 h post-intravenous administration using Lumina III Imaging System (PerkinElmer). The mice were sacrificed by cervical dislocation after last imaging. Tumors and major tissues were collected, fixed, dehydrated and sliced for fluorescence study48 (link). After stained the nuclei with DAPI (0.5% μg/mL), images of the slices were captured by a confocal microscope (Olympus).
3
Pharmacokinetics of d-SN38 Nanoparticles
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Female BALB/c mice (n = 3) were intravenously injected with d-SN38@NPs and d-SN38@NPs/iRGD at a dosage of 5 mg/kg of Ce6, respectively. 40 μL of blood sample was collected at 0.25, 0.5, 1, 2, 4, 8, 12, 24 and 48 h post injection. The Ce6 fluorescence signal (Ex 640 nm, Em 710 nm) of each sample was analyzed by Lumina III Imaging System (PerkinElmer, IVIS Lumina III, Waltham, MA, USA).
4
In Vivo Biodistribution of Theranostic Nanoparticles
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The model mice were vein‐intravenously injected at the dose of 0.5 mg kg−1 DiD (50 mg kg−1 TMNPs) with nanoparticles labeled by DiD (DiD@TMNPs and control nanoparticles). After injection for 1, 2, 4, 6, 8, and 12 h, the ASD mice were imaged using the Lumina III Imaging System (PerkinElmer, USA). At 12 h, the mice were sacrificed and their organs (heart, liver, spleen, lung, kidney, and brain) were separated, which were captured as above. All tissues were totally soaked into 4% paraformaldehyde for 24 h, then dehydrated with 10% and 30% sucrose solution for 24 h separately and embedded in Tissue‐Tek O.C.T compound (Sakura Finetek, USA). Then they were sectioned at 10 µm with the freezing microtome (Leica CM1950, Germany). Brain slides were operated by immunofluorescence of anti‐CD11b (bs‐1014R‐AF488, Bioss) antibody and others were stained with DAPI. The conditions of co‐localization were observed using a confocal microscope. All animals were maintained under SPF grade feeding conditions and experiments were approved by the Animal Experimentation Ethics Committee of Sichuan University (KS2020420).
5
Imaging of Nanoparticle Biodistribution in Alzheimer's Disease Mouse Model
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Firstly, AD model was established by injecting Aβ1–42 into hippocampus according to the method described previously38 (link). The male mice were anesthetized and fixed in a stereotactic device. Aβ1–42 was dissolved in artificial cerebrospinal fluid (1 mg/mL) and pre-incubated at 37 °C for 7 days. Next, Aβ1–42 (5 μL) was injected into the right hippocampus for 8 min and then withdrawn slowly. Two weeks after the operation, the mice were intravenously injected with the DiD-labeled nanoparticles and imaged by the Lumina III Imaging System (PerkinElmer, USA) at 2, 4, 8, 12, 24, 36 and 48 h post injection. At the same time, some mice were executed at the 8 h, and then their major organs were separated for ex vivo imaging. The fluorescence of the organs was measured as above. Next, the tissues were sectioned after the fixation and dehydration steps by freezing microtome (Leica CM1950, Germany). At last, the tissues were stained with DAPI (1 μg/mL). The images were observed using a confocal microscope.
6
Biodistribution of DiD-Labeled Nanoparticles
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4T1 cells (3 × 105 cells) were transplanted into the left mammary fat pad of female BALB/c mice. After 10 days, DiD@BNP and DiD@MBNP (0.3 mg DiD/kg) were intravenously injected into 4T1-bearing BALB/c mice. At 2, 6, 12 and 24 h, the fluorescence distribution was observed by the Lumina III Imaging System (PerkinElmer, USA). Twenty-four hours post injection, main organs were excised for imaging. Then, all the tumors and organs were dehydrated and frozenly sectioned to a 10 μm thickness with the freezing microtome (Leica CM1950, Germany). The DiD distributions were observed using a confocal microscope (A1R+, Nikon, Japan).
7
Biodistribution of Fluorescent Nanoparticles
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Tumor models were established and intravenously injected with DiD-NP, DiD-RNP, DiD-mRNP and DiD-ARNP via the tail vein. The biodistribution of DiD in the primary tumor and bone metastatic tumor was analyzed at 2, 6, 12 and 24 h after administration by applying the Lumina III Imaging System (PerkinElmer, USA). At the end of this experiment, all the mice were sacrificed. Major organs (heart, liver, spleen, lung, and kidney) and tumors were collected for ex vivo imaging of DiD fluorescence.
8
Tracking Nanoparticle Biodistribution in AD Mice
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The AD mice were intravenously injected with Cy5.5-Ibu-labeled nanoparticles (1 mg/kg of Cy5.5-Ibu, equal to 50 mg/kg of RNPs). After injected for 1, 2, 4 and 6 h, mice were imaged using the Lumina III Imaging System (PerkinElmer, USA). At 6 h, the mice were sacrificed and their organs (heart, liver, spleen, lung, kidney and brain) were separated and captured as above. All tissues were totally soaked into 4% paraformaldehyde for 24 h, dehydrated with sucrose solution step by step and embedded in Tissue-Tek O.C.T compound (Sakura Finetek, USA). Then they were sectioned at 10 μm with the freezing microtome (Leica CM1950, Germany). Brain slides were operated by immunofluorescence of anti-RAGE antibody and other organs were stained with DAPI. The images of co-localization were observed using a confocal microscope.
9
In Vivo Biodistribution of Nanoparticles
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Female BALB/c nude mice (∼20 g, n = 5) were orthotopically implanted with 4T1 cells at a density of 5 × 105 into the second breast pad on the left side. Then nanoparticles were intravenously injected with CAuNCs-GOx@HA, CAuNCs-Cat@HA and NM, respectively at a dosage of 1 mg BSA each. Major organs and tumors were collected and imaged by Lumina III Imaging System (Perkin Elmer, USA). And the fluorescence intensities were semi-quantified for comparison. Further, tissues were sliced and stained with DAPI, the fluorescence of nanoparticles were observed through confocal microscope (Nikon, A1R+, Japan).
10
In Vivo Bioluminescence Imaging of Cells
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Imaging was performed at the Vivoptic platform (University of Bordeaux, Bordeaux) using a Lumina III imaging system (Perkin Elmer Inc., Boston, MA, USA). HEK293T transfected cells were imaged (0.5 s) by bioluminescence 6 min after adding NanoGlo substrate (25 µM in 100 μL PBS; Promega).
For in vivo experiments and to determine the optimal experimental conditions, various NanoGlo substrate FFZ (Promega, Madison, WI, USA) quantities were injected intraperitoneally into the mice (0.35, 0.175, 0.088, and 0.044 µmoles). Three minutes after the substrate injection, mice were sedated, and bioluminescence acquisitions were performed (1 min, 4 × 4 binning) at 4, 6, 8, 10, and 12 min post-injection. Finally, mice received an intraperitoneal injection of 0.088 µmoles of NanoGlo substrate FFZ (Promega, Nano-Glo® Fluorofurimazine FFz) for all the bioluminescence acquisitions and were sedated 3 min later. Bioluminescence images and photographs (100 ms) were taken 6 min after the substrate injection. Image quantification was done using Living Image software (Perkin Elmer). At 10 weeks post-surgery, mice were euthanized by cervical dislocation, and organs were then rapidly retrieved for BLI.
For in vivo experiments and to determine the optimal experimental conditions, various NanoGlo substrate FFZ (Promega, Madison, WI, USA) quantities were injected intraperitoneally into the mice (0.35, 0.175, 0.088, and 0.044 µmoles). Three minutes after the substrate injection, mice were sedated, and bioluminescence acquisitions were performed (1 min, 4 × 4 binning) at 4, 6, 8, 10, and 12 min post-injection. Finally, mice received an intraperitoneal injection of 0.088 µmoles of NanoGlo substrate FFZ (Promega, Nano-Glo® Fluorofurimazine FFz) for all the bioluminescence acquisitions and were sedated 3 min later. Bioluminescence images and photographs (100 ms) were taken 6 min after the substrate injection. Image quantification was done using Living Image software (Perkin Elmer). At 10 weeks post-surgery, mice were euthanized by cervical dislocation, and organs were then rapidly retrieved for BLI.
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