Immunohistochemical staining was performed according to published methods [30 (link)]. First, 3 μm paraffin sections of tissue samples were stained with immunohistochemistry. The primary antibody specific for Ki-67 (Abcam, Cambridge, USA) was used at a 1:100 dilution in the experiments. Images were captured using a Nikon Eclipse 80i system with NIS-Elements software (Nikon, Japan).
Eclipse 80i system
Manufactured by Nikon
Sourced in Japan
The Nikon Eclipse 80i system is a high-performance research microscope designed for advanced imaging and analysis applications. It features a sturdy, ergonomic design and offers a range of advanced optical components, including a high-resolution objective lens system and a versatile illumination system, to facilitate detailed observation and documentation of samples.
Automatically generated - may contain errors
Lab products found in correlation
Nis elements software, by Nikon (3 mentions)
Ivis spectrum imaging system, by PerkinElmer (2 mentions)
Primary antibody specific for ki 67, by Abcam (1 mentions)
G3bp2, by Proteintech (1 mentions)
P src, by ABclonal (1 mentions)
P fak, by ABclonal (1 mentions)
E cadherin, by Proteintech (1 mentions)
N cadherin, by ABclonal (1 mentions)
Hmga2, by ABclonal (1 mentions)
Vimentin, by ABclonal (1 mentions)
Klf10, by Santa Cruz Biotechnology (1 mentions)
Matrigel, by BD (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
N cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
Transwell apparatus, by Corning (1 mentions)
Hematoxylin, by Maclin Biochemical Technology (1 mentions)
Matrigel, by R&D Systems (1 mentions)
13 protocols using eclipse 80i system
1
Immunohistochemical Staining of Ki-67
2
Examining Lymphatic Metastasis in Bladder Cancer
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The BALB/c nude mice (4–5 weeks old, 18–20 g) were purchased from the Experimental Animal Center, Sun Yat-sen University (Guangzhou, China). All experimental procedures were approved by the Institutional Animal Care and Use Committee of Sun Yat-sen University. The footpads of 16 mice were inoculated with 100 μl PBS suspensions of bladder cancer cells that were transduced with LNMAT1-luc, vector-luc, shLNMAT1-luc or sh-NC-luc. One week after the tumor cell inoculations, the nude mice were randomly selected for treatment by intraperitoneal injection with the PBS control or the CCL2 neutralizing antibody three times per week. Lymphatic metastasis was monitored and imaged with a bioluminescence imaging system (PerkinElmer, IVIS Spectrum Imaging System) 4 weeks after the injections. The tumor growth and that lymphatic metastasis were checked when the control tumors reached the same size as the experimental tumors. The primary tumors and popliteal LNs were enucleated and paraffin embedded. Serial 4.0-mm sections were obtained and analyzed by IHC. Images were captured using a Nikon Eclipse 80i system with NIS-Elements software (Nikon, Japan). The uncropped pictures are summarized in Supplementary Figure 15 .
3
Immunohistochemical Analysis of Signaling Proteins
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Immunohistochemical staining was performed according to published methods [5 (link)]. First, 5-μm paraffin sections of tissue samples were stained with HE and immunohistochemistry. The primary antibodies specific for G3BP2 (Proteintech, USA), SRC (ABclonal, China), p-SRC (ABclonal, China), FAK (ABclonal, China), and p-FAK (ABclonal, China) were used at a 1:100 dilution in the experiments. Images were captured using a Nikon Eclipse 80i system with NIS-Elements software (Nikon, Japan).
4
Investigating Tumor Growth Inhibition by hsa_circ_0006948
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Ethical approval was obtained from the Institutional Research Ethics Committee of Sun Yat-sen University. All animal care and procedures were performed in accordance with institutional guidelines. To assess the effect of hsa_circ_0006948 on tumor growth in xenograft models, TE-1 cells (5×106/0.2 ml PBS) were injected subcutaneously into the right backs of 4-week-old BALB/c nude mice. After 12 days, the mice were treated with intertumoral injection of negative control, si-hsa_circ_0006948 and combined si-hsa_circ_0006948 and agomiR-490-3p every two days, respectively. The tumors were measured every three days and their volumes were calculated according to the following formula: tumor volume = (length × width2)/2. Thirty days later, the mice were sacrificed, and the tumors were excised for further immunohistochemistry. Primary antibodies against E-cadherin (Proteintech, USA), vimentin (ABclonal, China), N-cadherin (ABclonal, China) and HMGA2 (ABclonal, China) were used. We captured images using a Nikon Eclipse 80i system with NIS-Elements software (Nikon, Japan).
5
Immunohistochemical Analysis of KLF10 and Slug
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This experiment was performed as previously described.41 (link) The paraffin-embedded tissues from nude mice were cut into 4-μm slides for H&E and IHC. The primary antibodies used in this study were KLF10 (Santa Cruz Biotechnology, 1:200) and Slug (Cell Signaling Technology, 1:200). The images were obtained by using a Nikon Eclipse 80i system (Nikon, Tokyo, Japan).
6
GADD45B Protein Expression in PCa
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GADD45B (1:100; YN1622; Immunoway; Beijing, China) antibody was used to assess the protein level in the PCa samples from Sun Yat-sen University Cancer Center via immunohistochemistry (IHC). IHC was performed according to standard procedures as described in our previously study (Wang et al., 2017 ). The immunoreactivity score (IRS) was calculated according to the following formula: IRS = intensity score × percentage score; intensity score: negative = 0, weak = 1, moderate = 2, and strong = 3; percentage score: <25% = 1, 25–50% = 2, 50–75% = 3, and >75% = 4. The samples were classified as low (IRS ≤ 6) or high (IRS > 6) GADD45B expression. The IRS score was blindly quantified by two pathologists. The photographs were taken using a Nikon Eclipse 80i system (Nikon, Tokyo, Japan).
7
Immunohistochemical Analysis of NEIL3 in PCa
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Ki67 (1:500; Servicebio; Wuhan, China) and NEIL3 antibodies (1:300; ab230908; Abcam, Cambridge, UK) were used to assess the protein level in PCa tissues and TMA were used for mouse tumors via immunohistochemistry (IHC) according to standard procedures. The images were acquired for statistical analysis using a Nikon Eclipse 80i system (Nikon, Tokyo, Japan). NEIL3 protein expression in the PCa samples was blindly quantified by 2 researchers. First, we evaluated the immunostaining intensity of each sample as follows: negative = 0, weak = 1, moderate = 2, and strong = 3. Second, we assessed the proportion of positively stained cells: < 25% = 1, 25%–50% = 2, 51%–75% = 3 and > 75% = 4. The immunoreactivity score (IRS) was calculated as the intensity score multiplied by the proportion score. The results were divided into 2 groups: low (IRS ≤ 6) and high (IRS > 6).
8
Immunohistochemical Analysis of KLF4
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Fresh tumor tissue samples from the nude mice were fixed in 4% paraformaldehyde, dehydrated in an ethanol solution, embedded in paraffin, and cut into 5 μm thick sections. The primary antibody for KLF4 (abcam, China) was used at a 1:1000 dilution in the experiments. Images were captured using a Nikon Eclipse 80i system with NIS-Elements software (Nikon, Japan).
9
Investigating CLDN6 in Cell Migration
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In order to reveal the effects of CLDN6 on cell migration and invasion, trans-well experiments were implemented using 6.5 mm transwell chambers with 8.0 µm pore-size polycarbonate membranes with or without Matrigel (BD Biosciences). In the invasion experiment, the upper chambers were first coated with 100 µL of Matrigel. In both invasion and migration assays, cells were suspended in serum-free medium at a 1:6 dilution (BD Biosciences) and incubated at 37°C for 4 hours. Subsequently, cells were loaded onto the top chamber of the transwell at a density of 5×105 cells/mL (200 µL/chamber). The lower chambers were filled with 500 µL complete media containing 10% FBS. After overnight incubation at 37°C in an air/5% CO2 atmosphere, non-invasive cells were removed from the top chambers with cotton swabs. The remaining cells that were attached to the underside of the membranes were fixed using 4% paraformaldehyde and stained with 0.1% crystal violet for 20 minutes. Next, the cells were counted in 5 randomly selected fields of view using a microscope (Nikon ECLIPSE 80i system), and the average value was recorded for every field. Each experiment was conducted at least 3 times.
10
Fluorescence-Based Bacterial Strain Characterization
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The pCm-GFP and pCm-GFP-RC were transformed into E. coli DH5α. For each strain, three colonies were randomly picked and inoculated respectively into 10 ml LB medium and incubated at 37 °C in a shaker at 200 rpm, while the background fluorescence was measured using DH5α harboring pCm. The fluorescence (excitation: 470 nm; emission: 510 nm) and OD600 were measured at 12 h. The fluorescence microscopy images were captured at 12, 18, and 24 h from drops of culture solution containing fluorescent cells by a Nikon Eclipse 80i system equipped with DS-Ri1 camera. The ratio of GFP to OD600 (GFP/OD600) was used to represent fluorescence intensity. The DH5α strain harboring plasmids pCPB-37-441 or pCPB-37-441-RC was inoculated into 10 ml LB medium containing 0.1 mM IPTG and incubated in a shaker at 200 rpm at 37 °C.
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