Tumor tissues were stained with anti-Ki67 antibody (CST, Danvers, MA, USA). Ki67-positive cells were observed and photographed with a Zeiss Axioskop 2 plus microscope. The proliferation index (PI) was quantified by determining the number of Ki67 positive cells among the total number of resting cells [28 (link)]. Cluster of differentiation 31 (CD31) is an important biomarker for vascular endothelial cells, and its immunostaining density is considered the tumor microvessel density (MVD) [29 (link)]. Tumor tissues were stained with anti-CD31 antibody (CST, Danvers, MA, USA) to detect the tumor MVD. Vessels with a clearly defined lumen or well-defined linear vessel shape were counted from the representative tumor zone using a Zeiss Axioskop 2 plus microscope. Then tumor tissues were stained with anti-cleaved PARP antibody (ProteinTech, Chicago, IL, USA) and anti-p-ERK1/2 antibody (CST, Danvers, MA, USA) respectively. Positive cells were observed and photographed with a Zeiss Axioskop 2 plus microscope. The percentage of positive staining cells was measured by determining the number of positive cells among the total number of cells.
Axioskop 2 plus microscope
Manufactured by Zeiss
Sourced in Germany, United States, Italy, Canada, Japan, Denmark, France
The Axioskop 2 plus microscope is a high-performance laboratory microscope designed for a variety of applications. It features a stable and ergonomic design with advanced optical components for delivering clear and detailed images. The microscope offers a wide range of magnification options and is capable of supporting various imaging techniques, including brightfield, darkfield, and phase contrast.
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Lab products found in correlation
Axiovision 4, by Zeiss (22 mentions)
Axiocam hrc digital camera, by Zeiss (11 mentions)
Axiocam mrc5 ccd camera, by Zeiss (10 mentions)
Axiocam hrc, by Zeiss (9 mentions)
Axiocam hrc camera, by Zeiss (9 mentions)
Vectashield, by Vector Laboratories (7 mentions)
Avidin biotin blocking kit, by Vector Laboratories (6 mentions)
Axiovision software, by Zeiss (6 mentions)
Matrigel, by BD (5 mentions)
Dab substrate, by Vector Laboratories (5 mentions)
Hematoxylin, by Merck Group (5 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (5 mentions)
Paraformaldehyde (pfa), by Merck Group (5 mentions)
Neutral buffered formalin, by Thermo Fisher Scientific (4 mentions)
Antigen unmasking solution, by Vector Laboratories (4 mentions)
Axiocam camera, by Zeiss (4 mentions)
Axiocam, by Zeiss (4 mentions)
Lsm image browser, by Zeiss (4 mentions)
Trypan blue solution, by Merck Group (4 mentions)
Zen 2010, by Zeiss (4 mentions)
303 protocols using axioskop 2 plus microscope
1
Evaluating Tumor Angiogenesis and Cell Proliferation
2
Quantifying Tumor Proliferation and Angiogenesis
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Tumour sections were stained with anti‐Ki67 antibody (CST, Danvers, MA, USA) and Ki67‐positive cells were observed under a Zeiss Axioskop 2 plus microscope. The proliferation index (PI) was calculated by the percentage of the Ki67 positive cells out of the total number of tumour cells.29 Cluster of differentiation 31 (CD31) has been identified as an ideal biomarker for vascular endothelial cells, and its immunostaining density is considered the tumour microvessel density (MVD).30 Then tumour sections were stained by IHC using CD31 antibody (CST, Danvers, MA, USA) to determine the tumour MVD. Stained vessels with a clearly defined lumen or well‐defined linear vessel shape were photographed using a Zeiss Axioskop 2 plus microscope and counted from the representative tumour zone.
3
Quantifying Tumor Cell Proliferation and Angiogenesis
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Tumor samples were stained with anti-Ki67 antibody (CST, Danvers, MA, USA). Ki67-positive tumor cells were observed with a Zeiss Axioskop 2 plus microscope. The proliferation index (PI) was determined as the ratio of the number of Ki67-positive cells to the total number of counted tumor cells [34 (link)]. Cluster of differentiation 31 (CD31) is a key biomarker for vascular endothelial cells, and its immunostaining density is represented by the tumor microvessel density (MVD) [35 (link)]. Tumor tissues were stained with CD31 antibody (CST, Danvers, MA, USA). Vessels were observed and counted by using a Zeiss Axioskop 2 plus microscope [31 (link)].
4
Analyzing Embryo Development with DAG2:GUS
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Analysis of dag2 and wild-type embryos was performed under an Axioskop 2 plus microscope (Zeiss).
The DAG2:GUS line is the one described in Gualberti et al. [8 (link)]. Histochemical staining and microscopic analysis were carried out according to Blazquez et al. [37 (link)]. Stained embryos (after washing in 70% ethanol) were analysed and photographed under an Axioskop 2 plus microscope (Zeiss).
The DAG2:GUS line is the one described in Gualberti et al. [8 (link)]. Histochemical staining and microscopic analysis were carried out according to Blazquez et al. [37 (link)]. Stained embryos (after washing in 70% ethanol) were analysed and photographed under an Axioskop 2 plus microscope (Zeiss).
5
Histopathological Analysis of Esophageal Specimens
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The esophageal specimens were dissected together with a portion of the stomach in order to localize the proximal and distal esophagus. We fixated the esophageal specimens in EAF (ethanol/acetic acid/formaldehyde/saline at 40:5:10:45 v/v), horizontally embedded in paraffin. Following the standard procedure, sagittal sections (at 2.0 μm) of paraffin blocks were stained with hematoxylin and eosin (H&E).
We used a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy, Oberkochen, Germany) and a Zeiss AxioCam HRc digital camera for histopathological analysis and digital microscopic images respectively. Microscopic images were processed using the AxioVision 4 software (Carl Zeiss Vision, Munich, Germany), and organ damage was assessed qualitatively by a pathologist [15 (link)].
We used a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy, Oberkochen, Germany) and a Zeiss AxioCam HRc digital camera for histopathological analysis and digital microscopic images respectively. Microscopic images were processed using the AxioVision 4 software (Carl Zeiss Vision, Munich, Germany), and organ damage was assessed qualitatively by a pathologist [15 (link)].
6
Histological Tissue Analysis with IHC
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Tissues were fixed in either 4% formalin or acidified formaldehyde (ethanol/acetic acid/formaldehyde/saline at 40:5:10:45 v/v). Sections were prepared at 2 µm thickness from the paraffin blocks and stained with haematoxylin and eosin (HE) according to standard procedures. For immunohistochemistry (IHC), 4 um-thick sections were made. Antibodies used in this study are listed in Supplementary Table 4 .
Antibody staining was revealed using either diaminobenzidine or 3-amino-9-ethylcarbazole chromogen. Images were captured using a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy) equipped with a Zeiss AxioCam HRc digital camera, and processed using AxioVision 4 software (Carl Zeiss Vision).
Antibody staining was revealed using either diaminobenzidine or 3-amino-9-ethylcarbazole chromogen. Images were captured using a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy) equipped with a Zeiss AxioCam HRc digital camera, and processed using AxioVision 4 software (Carl Zeiss Vision).
7
Cell Migration Assay Protocol
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1 × 105 cells were seeded into the upper chamber in serum-free RPMI 1640 medium uncoated or coated with Matrigel (BD Biosciences, San Jose, CA, USA). Then 500 µl corresponding medium containing 10% FBS was added into the lower chamber. After incubation for 24 h, the cells were scrubbed with a cotton tip swab. The cells on the bottom surface of the membrane were fixed with 4% paraformaldehyde for 20 min at 37 °C and stained with 0.1% crystal violet for 10 min at 37 °C. Cell number was counted with a Zeiss Axioskop 2 plus microscope (Carl Zeiss, Thornwood, NY, USA).
8
Quantitative Analysis of Vimentin in Tumor Tissue
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Lung and liver tissues were collected and fixed in formalin fixative and embedded in paraffin. The immunohistochemistry (IHC) of vimentin (DAKO, M0725, dilution: 1:4,000) was conducted on 4‐μm‐thick sections according to standard procedures. The stained slides were examined blindly by a pathologist, and the number of tumorous lesions (more than 10 cancer cells) was scored in each of the sections. The sections were reviewed with a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy, Jena, Germany), and images were captured with a Zeiss AxioCam HRc digital camera and processed with AxioVision 4 software (both from Carl Zeiss Vision, München, Germany).
9
Paraffin-Embedded Tissue Histology
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Tissues and organs were collected and fixed in EAF fixative (ethanol:acetic acid:formaldehyde:saline at 40:5:10:45 v:v) and embedded in paraffin. Sections were prepared at 2 µm thickness from the paraffin blocks and stained with haematoxylin and eosin according to standard procedures. The sections were reviewed with a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy) and images were captured with a Zeiss AxioCam HRc digital camera and processed with AxioVision 4 software (both from Carl Zeiss Vision).
10
Histological Analysis of Mouse Tissues
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Tissues were removed from the mice directly after euthanasia. They were fixed in formalin and embedded in paraffin. Two micrometer-thick sections were made and stained by hematoxylin and eosin (H&E) according to standard procedures. Four micrometer-thick sections were made and stained with antibodies anti-S100 (DakoCytomation; clone Z0311), keratin 1 (Covance; Clone PRB-165P), F4/80 (Serotec; Clone MCA497) or CD3 (Neomarkers; Clone RM-9107) according to standard immunohistochemistry (IHC) protocols. The sections were reviewed with a Zeiss Axioskop2 Plus microscope (Carl Zeiss Microscopy, Jena, Germany), and images were captured with a Zeiss AxioCam HRc digital camera and processed with AxioVision 4 software (both from Carl ZeissVision, München, Germany).
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