Normal female nude mice (4–6 weeks old, 18–20 g) were injected with PEGylated MGNRs@DMSSs (100 µL, [Au]=5.3 mg mL-1). At 24 hours post injection, mice were sacrificed and all major organs (heart, liver, spleen, lungs, and kidneys) were harvested. Tissues were then sectioned and examined using H&E staining. Slides were examined with a NIKON Eclipse Ci optical microscope (NIKON Digital Sight DS-FI2; Nikon Corporation, Tokyo, Japan).
Digital sight ds fi2
Manufactured by Nikon
Sourced in Japan
The Digital Sight DS-Fi2 is a digital microscope camera designed for laboratory use. It captures high-quality images and video of microscopic specimens. The device features a CMOS sensor and supports a variety of resolutions and frame rates. It connects to a computer via a USB interface for image acquisition and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ci, by Nikon (26 mentions)
Smz1270i, by Nikon (5 mentions)
Ms 222, by Merck Group (3 mentions)
Nis elements d analysis software version 3.2, by Nikon (3 mentions)
Smz800n, by Nikon (3 mentions)
Smz25, by Nikon (2 mentions)
Eclipse ni u light microscope, by Nikon (2 mentions)
Apochromat hr plan 0.5 w d 136 mm objective, by Nikon (2 mentions)
C apochromat 40 1.2 n a objective, by Zeiss (2 mentions)
Nis elements analysis d software, by Nikon (2 mentions)
Smz18, by Nikon (2 mentions)
Image pro plus 6, by Media Cybernetics (2 mentions)
Smz800, by Nikon (2 mentions)
Smz1500, by Nikon (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Hoechst, by Merck Group (1 mentions)
Live dead cell imaging kit, by Thermo Fisher Scientific (1 mentions)
Bx61 microscope, by Olympus (1 mentions)
Oil red o solution, by Wuhan Servicebio Technology (1 mentions)
Mab3328, by Merck Group (1 mentions)
82 protocols using digital sight ds fi2
1
Biodistribution of PEGylated MGNRs
2
Evaluating Lipid Vesicle Therapy for Jurkat Cell-Induced Paralysis in Mice
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The female NPG mice were supplied by Beijing Vitalstar Biotechnology Co., Ltd. (Beijing, China) and were fed under SPF condition. The female C57BL/6 mice and female BALB/c nu/nu mice were obtained from Peking University Health Science Center, China. All mouse studies adhered to the principles of care and use of laboratory animals and were approved by the Institutional Animal Care and Use Committee of Peking University, China.
The NPG mice were intravenously injected with 1 × 106 Jurkat cells. At 2 h post cell infusion, different DOX-loaded lipid vesicles (4 mg/kg, calculated as DOX) were injected through the tail vein, and 11 days after cell infusion, lipid vesicles were injected again at a 2 mg/kg DOX dose. The mice were sacrificed when they are paralyzed. Survival time was evaluated and organs were harvested at 1 month for tissue section analysis. Leg bones treated with decalcifying solution and hearts were frozen and fixed on poly lysine coated glass slides. After hematoxylin-eosin (H&E) staining, morphological changes were observed by microscope. NanoSPECT/CT images were detected by NIKON digital sight DS-FI2 (NIKON Inc., Minato, Tokyo, Japan).
The NPG mice were intravenously injected with 1 × 106 Jurkat cells. At 2 h post cell infusion, different DOX-loaded lipid vesicles (4 mg/kg, calculated as DOX) were injected through the tail vein, and 11 days after cell infusion, lipid vesicles were injected again at a 2 mg/kg DOX dose. The mice were sacrificed when they are paralyzed. Survival time was evaluated and organs were harvested at 1 month for tissue section analysis. Leg bones treated with decalcifying solution and hearts were frozen and fixed on poly lysine coated glass slides. After hematoxylin-eosin (H&E) staining, morphological changes were observed by microscope. NanoSPECT/CT images were detected by NIKON digital sight DS-FI2 (NIKON Inc., Minato, Tokyo, Japan).
3
Alizarin Red Staining of Mineralization
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Alizarin
red staining was used to measure mineralization. Cells were washed
3 times for 15 min at room temperature with PBS and fixed with 4%
formaldehyde. Then, the cells were washed with deionized water and
stained with 1 mL of 40 mM ARS per well for 30 min. Deionized water
was used to wash the cells prior to imaging and quantification. The
images of stained samples were collected with a Nikon SMZ1500 and
a Nikon Digital sight DS-Fi2 camera (Nikon, Japan). Subsequently,
stained cultures were quantified by destaining for 15 min at room
temperature using 10% (w/v) CPC in 10 mM sodium phosphate at pH 7.0.
The ARS concentration was measured by using the absorbance at 562
nm.
red staining was used to measure mineralization. Cells were washed
3 times for 15 min at room temperature with PBS and fixed with 4%
formaldehyde. Then, the cells were washed with deionized water and
stained with 1 mL of 40 mM ARS per well for 30 min. Deionized water
was used to wash the cells prior to imaging and quantification. The
images of stained samples were collected with a Nikon SMZ1500 and
a Nikon Digital sight DS-Fi2 camera (Nikon, Japan). Subsequently,
stained cultures were quantified by destaining for 15 min at room
temperature using 10% (w/v) CPC in 10 mM sodium phosphate at pH 7.0.
The ARS concentration was measured by using the absorbance at 562
nm.
4
Assessing Corneal Endothelial Viability
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A calcein AM assay (LIVE/DEAD Cell Imaging Kit; Invitrogen, Barcelona, Spain) was performed on a peeled-off endothelial of two cryopreserved corneas from each protocol, following the manufacturer’s instructions of the kit and counterstaining with Hoechst (Sigma, Barcelona, Spain). Endothelia were visualized with the Olympus BX61 microscope (Olympus España S.A., Barcelona, Spain), coupled with the digital camera Olympus DP70 (Olympus España S.A., Barcelona, Spain). The open software FIJI [43 (link)] was used for image merging.
A vital stain with trypan blue and alizarine red was used to determine the endothelial integrity of 10 TK-cryopreserved corneas. Based on the protocol of Taylor et al. [44 (link)], thawed corneas were rinsed twice with phosphate buffer saline (PBS), stained with 0.2% trypan blue (Gibco, Madrid, Spain) for 90 s, rinsed twice with PBS, stained with 0.2% alizarin red (pH 4.2; Panreac, Barcelona, Spain) for 90 s, and rinsed twice before their pictures were taken with the camera Nikon digital sight DS-Fi2 coupled to the stereoscopy Nikon SM7 745T.
A vital stain with trypan blue and alizarine red was used to determine the endothelial integrity of 10 TK-cryopreserved corneas. Based on the protocol of Taylor et al. [44 (link)], thawed corneas were rinsed twice with phosphate buffer saline (PBS), stained with 0.2% trypan blue (Gibco, Madrid, Spain) for 90 s, rinsed twice with PBS, stained with 0.2% alizarin red (pH 4.2; Panreac, Barcelona, Spain) for 90 s, and rinsed twice before their pictures were taken with the camera Nikon digital sight DS-Fi2 coupled to the stereoscopy Nikon SM7 745T.
5
Histological Analysis of Murine Liver
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Liver tissues from mice in each group were collected and fixed in 10% formaldehyde neutral buffer solution for 24 h at 4˚C, followed by washing with tap water, dehydration in alcohol and embedding in paraffin. Subsequently, the 5-µm thick transversal sections were obtained, deparaffinized, dehydrated in ethanol (50-100%) and cleared with xylene. The sections were stained using hematoxylin and eosin (H&E) solution for 5 min at room temperature (Wuhan Servicebio Technology Co., Ltd.), according to the manufacturer's protocols. For Oil-red O staining, liver tissues were frozen in optimal cutting temperature compound at -20˚C and 10-µm thick sections were obtained, followed by staining with Oil-red O solution (Wuhan Servicebio Technology Co., Ltd.) for 10 min at room temperature, according to the manufacturer's protocols. Images were observed using a NIKON imaging workstation (NIKON digital sight DS-FI2; Nikon Corporation).
6
Chicken Embryo Xenograft Assay for Tumor Analysis
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Fertilized white Leghorn chicken eggs were incubated at 37 °C under constant humidity (embryo development day 0, EDD0). No method of randomization was used. Separation of the developing CAM was induced on EDD4. On EDD8, transfected PC3 cells were suspended in PBS and Matrigel (1:1) and implanted on the CAM. On EDD11, the tumors were photographed in ovo and excised. Tumor area was measured in blind on photographs from four independent experiments, each with 3–5 eggs per shRNA construct, using ImageJ (NIH, Bethesda, MD, USA). Tumor specimens were placed in 4% paraformaldehyde, paraffin embedded and cut in 5 μm sections. Sections were deparaffinized and stained with HE. For immunohistochemistry, antigen retrieval was carried out with 10 mm sodium citrate buffer (pH 6.0). Nonspecific binding was blocked with bovine serum albumin and normal goat serum, and sections were stained with anti-MMP14 primary antibody (MAB3328; Millipore, Darmstadt, Germany) overnight at 4 °C. Sections were pretreated with 0.3% hydrogen peroxide and incubated with horse radish peroxidase-conjugated secondary antibodies (Promega, Madison, WI, USA or GE Healthcare, Buckinghamshire, England), followed by incubation with 3,3′-diaminobenzidine chromogenic substrate. Digital images were captured with Leica DM RXA light microscope (Wetzlar, Germany) and Nikon Digital Sight DS-Fi2 (Tokyo, Japan).
7
Histopathological Analysis of Lung Tissues
8
Quantitative Analysis of Cartilage Proteoglycans
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After synchrotron microCT imaging, the frozen samples were thawed and cut in half. The first half was fixed in 10% formalin and then dehydrated in ascending series of ethanol for the reference histological analysis. After dehydration, the samples were decalcified in EDTA and embedded in paraffin in order to cut 3 µm thick sections. Then, the paraffin was removed, and Safranin-O staining was performed to study the spatial FCD (i.e. PG) distribution in the cartilage. After staining, quantitative digital densitometry measurements were conducted to study the optical density (OD, e.g., PG distribution) in cartilage using a light microscope (Nikon Microphot-FXA, Nikon Co., Japan) equipped with a monochromatic light source and a 12-bit CCD camera (ORCA-ER, Hamamatsu Photonics K.K., Japan). System was calibrated using neutral density filters (Schott, Germany) covering OD range from 0 to 2.6. Histological images of Safranin-O stained sections were also imaged with a light microscope (Leica MZ75, Leica Microsystems Ltd., Switzerland) fitted with a CCD camera (Nikon digital sight DS-Fi2, Nikon Co., Japan). The depth-wise PG content within each cartilage sample was determined as an average of three sections. From the second half, water content was determined by calculating the difference between the wet weight and dry weight after freeze-dying the samples for 20 h.
9
Ovarian Follicle Histological Analysis
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The histological variations in the follicles from the ovarian tissue were observed by H&E staining. First, the ovaries were embedded in paraffin blocks for sectioning (5 μm thick), then rehydrated for hematoxylin staining (3 min) and rinsed with tap water (15 min). The sections were then differentially stained using 1% hydrochloric acid–ethanol solution (15 s), washed with tap water (5 min), counterstained with eosin (5 min), washed with tap water (15 min), and sealed with neutral gum. Finally, the stained sections were imaged using an upright optical microscope (NIKON Eclipse ci, NIKON digital sight DS-FI2, Nikon, Japan). The ovarian structures were analyzed, according to the type of follicles: growing follicles with more than three layers of cubic granulosa cells and no antrum present [37 (link)]; antral follicles with multiple layers of granulosa cells and antrum present [38 (link)]; and atretic follicles, the morphology of which is complex (at least five pyknotic granulosa nuclei and/or degenerated oocytes; a highly vascularized luteinized cell surrounded by the pseudotheca, 210–430 μm in diameter; a highly vascularized luteinized cell capsule, 150–270 μm in diameter) [38 (link),39 (link)]. Three sections per ovary were examined.
10
Histopathological Evaluation of Mouse Liver
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Mouse liver tissues were harvested and immediately fixed in 4% methyl aldehyde, embedded in paraffin, cut into sections, and stained with hematoxylin and eosin (H&E) following a standard protocol. H&E-stained liver sections were used to evaluate liver damage using a Nikon digital sight DS-FI2 (Nikon, Japan). Moreover, Pannoramic slice scanner with CaseViewer 2.2 scanning and browsing software was used to select the equal target area of the slices. After imaging, Image-Pro Plus 6.0 software was used to measure the necrotic tissue area of 6 visual fields of each slice and the corresponding area of whole tissue in each slice. The percentage of necrotic tissue area was then calculated.
Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and the levels of total liver GSH were measured with a commercial reagent kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's instruction.
Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and the levels of total liver GSH were measured with a commercial reagent kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's instruction.
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