Haematoxylin‒eosin (H&E) staining was used to examine the histopathology of mouse lung tissues (Cat# ZLI‐9610, ZSGB‐BIO), and the results were quantified by a semiquantitative scoring system reported by Szapiel. Masson's trichrome staining (Cat# G1340, Solarbio Life Science) was used to detect collagen content in lung tissues, and collagen quantification was performed using ImageJ software.
Zli 9610
Manufactured by ZSGB-BIO
Sourced in China
The ZLI-9610 is a laboratory centrifuge designed for general-purpose use. It features a compact and durable construction, accommodating a range of sample volumes and tube sizes. The centrifuge is capable of reaching speeds up to 6,000 rpm, enabling effective separation of biological samples. Its simple and intuitive control panel allows for easy operation.
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Lab products found in correlation
Zli 9613, by ZSGB-BIO (5 mentions)
Matrigel matrix, by BD (3 mentions)
Upper transwell chamber, by BD (2 mentions)
Zli 9017, by ZSGB-BIO (2 mentions)
Masson s trichrome staining, by Solarbio (1 mentions)
Hematoxylin, by ZSGB-BIO (1 mentions)
Sap 9100, by ZSGB-BIO (1 mentions)
3 3 diaminobenzidine (dab), by ZSGB-BIO (1 mentions)
Anti ki67, by ZSGB-BIO (1 mentions)
Za 0502, by ZSGB-BIO (1 mentions)
Ab52490, by Abcam (1 mentions)
Zm 0166, by ZSGB-BIO (1 mentions)
Pv 9000, by ZSGB-BIO (1 mentions)
Ab64693, by Abcam (1 mentions)
Ab183685, by Abcam (1 mentions)
Ds 0004, by ZSGB-BIO (1 mentions)
Naphthol as mx, by Merck Group (1 mentions)
Fast red violet lb salt, by Merck Group (1 mentions)
Ep1863y, by Abcam (1 mentions)
G1100, by Solarbio (1 mentions)
16 protocols using zli 9610
1
Histopathological Lung Tissue Analysis
2
Immunohistochemical Analysis of CCL20 Expression
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The tumor tissues of mice were fixed in formalin and processed for paraffin embedding. Sectioned samples were deparaffined in xylene and rehydrated in graded alcohol. Antigen retrieval was done according to the manufacture’s protocol (MVS-0100, Maxvision), and then the endogenous peroxidase was inactivated with 3% hydrogen peroxide methanol solution, blocked with animal nonimmune serum (SP KIT-B, Maxvision) and incubated with primary antibodies overnight at 4°C, and then incubated with secondary antibodies for 15 minutes. Slides were stained using the detection kit (DAB-0031, Maxvision); cell nucleus was stained with hematoxylin (ZLI-9610, ZSGB-BIO). The following antibodies and dilutions were used for IHC: CCL20 (1:50, Cat AF360, R&D), peroxidase-conjugated anti-goat secondary antibody (KIT 5107, Maxvision). Images were captured by trinocular microscope (OLYMPUS BX43, Japan). CCL20 expression was scored semiquantitatively by a manual histo-score (H-score) methodology based on staining intensity and percentage of positive tumor cells. Strongly staining scored 3, moderately staining scored 2, weakly staining scored 1, and negatively staining scored 0. The H-score of CCL20 expression is obtained by the formula 3 × percentage of strongly staining + 2 × percentage of moderately staining + percentage of weakly staining, giving a range of 0–300.
3
Transwell Invasion and Migration Assay
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The cells were resuspended in serum-free medium the day before experiments. For Transwell invasion assays, Transwell upper chambers (8 mm; BD Biosciences, Franklin Lakes, BJ, USA) were coated with Matrigel matrix (dilution 1:7; 356234; BD Biosciences, Franklin Lakes, BJ, USA) and 2 × 105 cells were placed in. For Transwell migration assays, 2 × 104 cells were seeded in the upper chamber of the Transwell without Matrigel matrix. Then, a medium containing 20% serum was added to lower chambers. Cells were incubated at 37 °C and 5% CO2 for 18 h, and then those cells remaining in upper chambers were removed using a wet cotton swab. We used 4% paraformaldehyde to fix the cells that had migrated and adhered to lower chambers for 10 min. Then hematoxylin (ZLI-9610, ZSGB-Bio) and eosin (ZLI-9613, ZSGB-Bio) were used to stain cells adhered to lower chambers for 20 min and then imaged. The number of cells was counted in ten separated high power fields with vertical cross distribution.
4
Histological Analysis of Mouse Testes
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Mouse testes were fixed in 4% paraformaldehyde at 4 °C overnight and rinsed in 0.1 mol/L phosphate buffer for 10 min. Next, the testicles were progressively dehydrated using an ethanol gradient, penetrated with xylene, and embedded in paraffin. Sections 5 μm-thick were mounted on slides coated with L-lysine followed by deparaffinisation and rehydration. For haematoxylin and eosin (HE) staining, the slides were stained with haematoxylin (ZLI-9610, ZSbio, Beijing, China) and eosin (ZLI-9613, ZSbio), as described previously.44 (link)
5
Immunohistochemical Analysis of Brain Tissues
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The slices (paraffin sections) of normal brain tissues and tumour tissues were used for immunohistochemistry analysis. Briefly, the slices were dewaxed, gradient dehydrated with alcohol, washed with standard method, repaired with water bath in antigen repairing solution and cooled with tap water. Then, the slices were blocked by normal goat serum solution (SAP-9100, ZSGB-BIO, Beijing, China). Then, the slices were dealt with the anti-IL31RA (K008474P, Solarbio, Beijing, China) and goat anti-rabbit IgG, and the slices were washed with standard methods, respectively. After that, the slices were dripped with Streptomyces ovalbumin protein labelled with horseradish peroxidase, and the slices were color developed with DAB (ZLI-9017, ZSGB-BIO, Beijing, China). Hematoxylin (ZLI-9610, ZSGB-BIO, Beijing, China) was used for counterstaining. Finally, the pathologist observed and interpreted the stained tissues under a light microscope.
6
Immunohistochemical Analysis of ANXA3 and Ki67
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7
Immunohistochemical Evaluation of MSN Expression in Breast Cancer
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The tumor tissues of mice were fixed in formalin and processed for paraffin embedding. Sectioned samples were deparaffined in xylene and rehydrated in graded alcohol. Antigen retrieval was done according to the manufacturer’s protocol (MVS-0100, Maxvision), and then the endogenous peroxidase was inactivated with 3% hydrogen peroxide methanol solution, blocked with animal nonimmune serum (SP KIT-B, Maxvision), and incubated with primary antibodies overnight at 4°C and then with secondary antibodies for 15 min. Slides were stained using the detection kit (DAB-0031, Maxvision); cell nucleus was stained with hematoxylin (ZLI-9610, ZSGB-BIO). The following antibodies and dilutions were used for immunohistochemistry (IHC): MSN (1:100, ab52490, Abcam), Ki67 (1:100, ZM-0166, ZSGB-BIO), NONO (1:100, 11058-1-AP, Proteintech), and peroxidase-conjugated secondary antibody (KIT-5010, Maxvision). MSN expression in breast cancer tissue slices was scored semiquantitatively by a manual histo-score (H-score) methodology based on staining intensity and percentage of positive tumor cells. Strongly staining scored 3, moderately staining scored 2, weakly staining scored 1, and negatively staining scored 0. The H-score of MSN expression is obtained by the formula 3 × percentage of strongly staining + 2 × percentage of moderately staining + percentage of weakly staining, giving a range of 0 to 300.
8
Transwell Assays for Cell Migration and Invasion
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For Transwell migration assays, 2 × 104 cells were resuspended in a serum-free medium and seeded in the upper chamber of the Transwell (8 mm, BD Biosciences, Franklin Lakes, NJ, USA). Medium containing 20% serum was added into the lower chamber. After 18 hr of incubation at 37°C and 5% CO2, the cells remaining in the upper chamber were removed using a wet cotton swab. Transwell invasion assays were performed as described for the migration assays, except that the Transwell upper chamber was coated with Matrigel matrix (dilution 1:7; 356234, BD Biosciences), and 2 × 105 cells were placed in the upper chamber. The cells that had migrated and adhered to the lower chamber were fixed with 4% paraformaldehyde for 10 min, stained with hematoxylin (ZLI-9610, ZSGB-Bio) and eosin (ZLI-9613, ZSGB-Bio) for 20 min, and then imaged. The number of cells was counted in ten separate high-power fields with vertical cross distribution.
9
Immunohistochemical Analysis of GBM
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Tissue sections of animal tumor and 40 clinical GBM pathological sections from the Department of Pathology of Xiangya Hospital (2019–2021) were collected. IHC was performed using a universal two‐step detection kit (PV‐9000, ZSGB‐Bio) and a DAB kit (ZLI‐9017, ZSGB‐Bio). Then the sections were lightly counterstained with Mayer hematoxylin (ZLI‐9610, ZSGB‐Bio). The expression of each protein was semi‐quantitatively evaluated using the method previously described.14
10
Immunohistochemical Analysis of CD4 and CD206
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For Immunohistochemical analysis, the tissues were deparaffinized in xylene and rehydrated in gradient ethanol. After immersion in ethylenediaminetetraacetic acid (EDTA) antigenic retrieval solution (pH8.0), the tissue sections were incubated with the normal goat serum blocking buffer at room temperature for 1 h. The sections were subsequently probed to the anti-CD4 primary antibody (ab183685, 1: 200, Abcam) or anti-CD206 (ab64693, 1: 100, Abcam) at 4 °C overnight, and then incubated at room temperature for 1 h with peroxidase-conjugated secondary antibody (DS-0004, ZSGB-BIO, Beijing, China). The cell nuclei were subjected to staining with the hematoxylin (ZLI-9610, ZSGB-BIO).
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