Brightfield images of frozen tissue sections were collected using a TissueFAXS PLUS system (TissueGnostics, Tarzana, CA) and analyzed in Adobe Photoshop (Adobe, San Jose, CA). Dermal cells/mm2 were consistently counted at 0.24 mm below the epidermal basal layer across all samples, over the width of each tissue section. To evaluate epidermal staining for proliferating cells, we divided the number of stained cells by the area occupied by the epidermis. Epidermal thickness was measured by dividing the epidermal area by the epidermal length. Where a percentage of stained cells was evaluated to measure T cell subset abundance, transcription factor staining was quantified among at least 200 CD3+ cells if available. Multispectral images were collected as whole slide scans using the Vectra 3 Automated Quantitative Pathology Imaging System (Akoya Biosciences). Spectral unmixing, tissue segmentation, and phenotyping were performed using inForm tissue analysis software (version 2.4.10, Akoya Biosciences). Quantification of cell density, marker co-expression, and colocalization were performed on whole slide scans using phenoptrReports software (Akoya Biosciences) (Viratham Pulsawatdi et al., 2020 (link)).
Tissuefaxs plus system
Manufactured by TissueGnostics
Sourced in Austria, Germany
The TissueFAXS PLUS system is a high-performance microscopic imaging platform designed for the acquisition and analysis of high-resolution digital images of tissue samples. The system utilizes advanced camera and illumination technologies to capture detailed images of biological specimens, enabling comprehensive visual analysis and quantification of various cellular and tissue parameters.
Automatically generated - may contain errors
Lab products found in correlation
3 3 diaminobenzidine tetrahydrochloride dab, by Agilent Technologies (2 mentions)
Axio imager 2 microscope, by Zeiss (2 mentions)
Strataquest analysis software, by TissueGnostics (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Vectra 3 automated quantitative pathology imaging system, by Akoya Biosciences (1 mentions)
Inform tissue analysis software, by Akoya Biosciences (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Novus Biologicals (1 mentions)
Histoquest, by TissueGnostics (1 mentions)
Permount, by Merck Group (1 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
Observer microscope, by Zeiss (1 mentions)
Histoquest software, by TissueGnostics (1 mentions)
Sk 4100, by Vector Laboratories (1 mentions)
Pk 6100, by Vector Laboratories (1 mentions)
Colibri 7 light source, by Zeiss (1 mentions)
Hematoxylin eosin he, by Merck Group (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Plank rychlo s solution, by Muto Pure Chemicals (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
3 3 diaminobenzidine substrate, by Vector Laboratories (1 mentions)
15 protocols using tissuefaxs plus system
1
Quantitative Multiplex Tissue Imaging
2
Immunohistochemical Analysis of Tumor Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor tissues were fixed in 4% polyoxymethylene, embedded in paraffin, sectioned to 3-μm pieces, and mounted on adhesion microscope slides. Deparaffinization and rehydration were performed in the following standard procedure. Antigen retrieval was performed by boiling slides in EDTA buffer (pH 9) for 20 minutes, and the slides were cooled on bench top for 20 minutes. The activity of endogenous peroxidases was quenched by incubation in 3% H2O2 for 10 minutes at room temperature. After being washed with PBS three times, the slides were then incubated with primary antibodies at 4°C overnight. The slides were equilibrated to room temperature for 1 hour. Slides were washed with PBS three times for 5 minutes. Secondary antibodies were added to slides and incubated for 1 hour. After being washed three times, slides were visualized using diaminobenzidine as a chromogenic substrate for 2 minutes and counterstained with hematoxylin for 2 minutes. Differentiation was performed using 1% HCl in 75% ethanol for 30 seconds. Quantitative analysis of ligand expression levels between tumor and nontumor cells was performed by TissueGnostics Asia Pacific Limited. The slides were scanned by a TissueFAXS Plus system (TissueGnostics GmbH) and analyzed by StrataQuest.
3
Immunohistochemical Quantification of Collagen I and OPG
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After dewaxing thoroughly with xylene and ethanol, the endogenous peroxidase activity was quenched with 3% hydrogen peroxide for 15 min. Antigen retrieval was then performed with citrate buffer at 80 °C and pH 6.5 for 10 min for immunohistochemistry detection. The tissue sections were reacted and incubated in a 1:100-diluted anti-collagen I antibody solution and a 1:100 diluted anti-osteoprotegerin (OPG) solution (Novus Biologicals, Littleton, CO, USA) at 37 °C for 1 h. The specimens were further incubated in a horseradish peroxidase-conjugated secondary antibody (Novus Biologicals, Littleton, CO, USA) for 30 min at room temperature. Finally, the sections were counterstained with hematoxylin after developing with 3,3′-diaminobenzidine tetrahydrochloride (DAB) (DAKO, Glostrup, Denmark), resulting in a brown color [50 (link),51 (link)]. Histologic observations and images were acquired using a TissueFAXS plus system (TissueGnostics GmbH, Vienna, Austria) under ×10 magnification. The DAB total area / tissue total area (%) was determined using HistoQuest (TissueGnostics, Los Angeles, CA, USA) analysis software.
4
Colocalization of Chemokines in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The colocalization of CCL11 and CCR3 was confirmed using confocal microscopy. The immunofluorescence of CCL11 overexpression FaDu and NPC204 cells was studied using CCL11 and CCR3 primary antibodies. The cells were incubated, respectively, with fluorescent secondary antibodies Alexa Fluor® 488 or 647 (Thermo Fisher Scientific, Waltham, MA, USA), followed by a nuclear counterstaining with DAPI. Confocal images were visualized at 20× magnification using a TissueFAXS Plus System (TissueGnostics, Vienna, Austria) coupled onto a Zeiss® Axio Imager Z2 microscope (Jena, Germany). Annotated cell borders were identified with the transmission channel.
5
Slit2 Protein Expression in SHR Striatum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the expression of the Slit2 protein in the striatum of SHR, immunohistochemistry (IHC) was performed as described elsewhere [37 (link)]. Animals were euthanized by carbon dioxide. The striatum tissues were excised, soaked in 10% formalin, and subsequently embedded with paraffin wax. The embedded tissues were sectioned into 5 μm slices and incubated overnight with antibodies against rat Slit2 (Cat. #: ab7665, Abcam, Waltham, MA, USA). Finally, the sections were observed and quantified using the automated Tissue-FAXS PLUS system (TISSUE GNOSTICS, Vienna, Austria).
6
Histological Analysis of Tail Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tail tissue blocks were cut into 5-μm sections. For general histological examination, deparaffinized sections were stained with hematoxylin and eosin (H&E). The distance of the distance between the epithelial migration tongues (dMT) or the distance between the hair follicles (dA) was determined as the descriptions from Gerharz et al. [24 (link)] using ImageJ software (Version 1.53 m, National Institutes of Health, Bethesda, MA, USA). The ratio of re-epithelialization was calculated by the formula of dMT/dA. For immunohistochemical staining, the deparaffinized tail sections were reacted with 3% H2O2 for 20 min. After blocking with 2% bovine serum albumin for 1 h at 25 °C, samples were incubated with mouse monoclonal anti-human MMP1 antibody (1:100, Cat. No. MAB3307, EMD Milli-pore Corporation) and mouse monoclonal anti-αSMA antibody (1:100, Cat. No. sc-32251, Santa Cruz Biotechnology, Inc.) overnight at 4 °C and then with the corresponding secondary antibody for 1 h at 37 °C. Antigenic sites were visualized by administering 3,3′-diaminobenzidine and hematoxylin for counterstaining. All slides were scanned through a TissueFAXS PLUS System (TissueGnostics, Taborstrasse, Vienna, Austria).
7
Immunohistochemical Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin sections (5 μm) were deparaffinized and incubated with the primary antibodies (1:100 dilution) using a standard avidin–biotin–peroxidase complex method. The slides were incubated with 3,3′-diaminobenzidine (DAKO, Carpinteria, CA, USA) to detect the antibody binding. The slides were then counterstained with hematoxylin, mounted with Permount (Merck, Darmstadt, Germany), and scanned/analyzed with a TissueFAXS PLUS system (TissueGnostics, Vienna, Austria).
8
Analyzing G9a and Hematoxylin Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The slides were digitally scanned using the TissueFAXS PLUS system (version 4.2.6245.1019, TissueGnostics, Vienna, Austria) on a Zeiss Observer microscope and the HistoQuest software (version 4.0.4.0158, TissueGnostics, Vienna, Austria) was used to analyze the G9a and hematoxylin staining and cell counts. Image analysis was performed in different regions of interest (ROI) in each image slide. G9a positive cell expression was calculated in percentages throughout the ROIs, and at least three representative areas were measured. The pixels were converted to grayscale and assigned an arbitrary number relating to the staining intensity, and then values were optimized by counting the number of DAB-positive and hematoxylin-positive events.
9
Immunohistochemical Analysis of Ovary Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ovaries were fixed in 4% paraformaldehyde and embedded in paraffin. The sections were stained with immunohistochemistry using the method as reported [34 (link)]. Ovary sections were incubated with primary antibody, reacted with biotin-labeled secondary antibodies (Vector Laboratories catalog no. PK-6100, RRID:AB_2336819), and stained with a 3, 3V-diaminobenzidine peroxidase substrate kit (Vector Laboratories catalog no. SK-4100, RRID:AB_2336382). Sections were counterstained with hematoxylin and analyzed with the TissueFAXS Plus system (Tissue Gnostics, Austria). Information on the relevant primary antibody is listed in Table 1 .
10
Fluorescence Imaging Protocol for Cell Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescence imaging was conducted using an Axio Imager 2 microscope (20×, ZEISS, Jena, Germany) equipped with a Colibri 7 light source (ZEISS, Germany) and the TissueFAXS PLUS system (TissueGnostics, Vienna, Austria). The light source comprises six LED modules and seven fluorescence channels, each emitting monochromatic light of a different wavelength. LED-optimized filters and direct coupling enhance sensitivity and ensure optimal excitation and emission spectra.
The images were analyzed using StrataQuest Analysis Software (v7, TissueGnostics, Vienna, Austria), and the minimum and maximum ranges for each filter were set by automatically adjusting the saturation. The mean minimum and maximum intensities of the slides were used for each marker in each panel [22 (link)].
Nuclei were detected and segmented using DAPI images. The mean staining intensities for the five different markers were measured using nuclei areas in six equally distributed circular ROIs with an area of 0.785 mm2 (a circular ROI that fits into a 1 × 1 cm square) per slide. The total number of cells with a mean intensity greater than 100, which were considered as ‘positive’, was recorded.
The images were analyzed using StrataQuest Analysis Software (v7, TissueGnostics, Vienna, Austria), and the minimum and maximum ranges for each filter were set by automatically adjusting the saturation. The mean minimum and maximum intensities of the slides were used for each marker in each panel [22 (link)].
Nuclei were detected and segmented using DAPI images. The mean staining intensities for the five different markers were measured using nuclei areas in six equally distributed circular ROIs with an area of 0.785 mm2 (a circular ROI that fits into a 1 × 1 cm square) per slide. The total number of cells with a mean intensity greater than 100, which were considered as ‘positive’, was recorded.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!