Phosphate buffered saline, buffered 10% formalin, Triton-X 100 Tween 20, tri-sodium citrate dehydrate and acid alcohol were purchased from Sigma-Aldrich (Steinheim, Germany), ethanol (HmbG Chemicals, Germany), xylene (Fisher Scientific, USA), hematoxylin and eosin (BDH, Germany), DPX (R&M Chemicals, UK), absolute propylene glycol solution and glycerin jelly M-12 (Rowley Biochemical Inc., USA), Peroxidazed 1 (BioCare Medical, USA), Blocking One (Histo Nacalai Tesque, Inc, Japan), ImmunoCruz ABC Staining and Ultra-Cruz Mounting Media were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All other chemicals used were of analytical grade.
Acid alcohol
Manufactured by Merck Group
Sourced in Germany
Acid alcohol is a laboratory reagent used to dissolve or dilute certain substances. It is a mixture of alcohol, typically ethanol or methanol, and an acid, such as hydrochloric acid or acetic acid. The specific composition may vary depending on the intended application. Acid alcohol is commonly used in various analytical and preparatory procedures in scientific research and clinical settings.
Automatically generated - may contain errors
Lab products found in correlation
Xylene substitute, by Thermo Fisher Scientific (7 mentions)
Permount, by Thermo Fisher Scientific (3 mentions)
Autostainer xl, by Leica (2 mentions)
Hematoxylin, by Merck Group (2 mentions)
Matrigel, by Corning (2 mentions)
Trisodium citrate dehydrate, by Merck Group (1 mentions)
Hematoxylin and eosin, by BD (1 mentions)
Xylene, by Thermo Fisher Scientific (1 mentions)
Peroxidazed 1, by Biocare Medical (1 mentions)
Ultra cruz mounting media, by Santa Cruz Biotechnology (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Harris haematoxylin, by Merck Group (1 mentions)
Faramount aqueous mounting medium, by Agilent Technologies (1 mentions)
Scott s tap water, by Merck Group (1 mentions)
8 protocols using acid alcohol
1
Tissue Preparation for Histological Analysis
2
Histological Analysis of Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were sacrificed and the subcutaneous tumors were dissected. Tumors were fixed in 10% formalin (Fisher, cat. SF98–4) at room temperature overnight and consequentially dehydrated in 70% ethanol. Samples were mounted in paraffin blocks and sectioned at 5 μm of thickness. Haematoxylin and eosin staining was performed in Lecia Autostainer XL. Briefly, slides were baked at 60°C for 15 minutes, then treated with Xylene Substitute (Citrisolv) (Fisher chemical cat. x3p-1gal) then rehydrated and stained with Hematoxylin, followed with washes with running water. An incubation with Acid Alcohol (Sigma, cat. A3429) was performed, followed with washes with running water. The samples were then stained with eosin (Thermo Scientific) then dehydrated with ethanol followed by Xylene Substitute (Citrisolv). Samples were mounted with Permount (Fisher Scientific, cat. SP15–500). Slides were submitted for pathologic evaluation. Pathologist was blinded to sample origin (i.e., experimental versus control)
3
Transwell Migration Assay for PDAC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 4 × 104 PDAC cells were plated on a 24-well Transwell polycarbonate membrane with 8.0-μm pore size (Corning 3422). Wells were coated with 50 μl of 3 mg/ml Matrigel (Corning 354230) and kept at 37°C for 24 hours before plating to ensure solidification. Cells plated in the upper chamber, unless treated with conditioned media, were placed in 150 μl of the appropriate serum-free media, whereas the bottom chamber contained 700 μl of the appropriate serum-containing media to act as the chemoattractant in the study. Cells were collected at 48 hours and gently rinsed with ddH2O and then fixed for 20 minutes with 10% formalin (Azer Scientific). After fixation, the cells were placed in hematoxylin (Sigma) for 2 minutes, rinsed in ddH2O, quickly dipped in 1% acid alcohol (Sigma), and rinsed a final time in ddH2O. Membranes were then imaged, and individual cells were counted manually using an AMG EVOS XL Core Cell Imaging System microscope (AMEX1000).
4
Transwell Assay for Cell Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
40,000 epithelial cells (25,000 cancer-associated fibroblasts) were plated on a 24-well transwell polycarbonate membrane with 8.0 μm pore size (Corning 3422). Wells used for migratory studies were coated with 50 μL of 3 mg/mL Matrigel (Corning 354230) and kept at 37°C for 24 hours before plating to ensure solidification. Cells plated in the upper chamber, unless treated with conditioned media, were placed in 150 μL of the appropriate serum-free media, whereas the bottom chamber contained 700 μL of the appropriate serum-containing media to act as the chemoattractant in the study. Once the cells were plated, the assay was run for 48 hours before cells were briefly and gently rinsed with ddH2O and then fixed for 20 minutes with 10% formalin (Azer Scientific). After fixation, the cells were placed in hematoxylin (Sigma) for 2 minutes, quickly rinsed in ddH2O, quickly dipped in 1% acid alcohol (Sigma), and rinsed a final time in ddH2O. Membranes were then imaged and individual cells were counted manually using an AMG EVOS XL Core Cell Imaging System microscope (AMEX1000).
5
Histological Analysis of Tumor Samples
6
Quantifying Lung Metastasis in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were sacrificed at four weeks (Experiment NO 1 and 2 in vivo) after secondary injection and the lung tissue was dissected. The tissue was fixed in 10% formalin at room temperature overnight and consequentially dehydrated in 70% ethanol. Samples were mounted in paraffin blocks and sectioned at 5 mm of thickness. Slides were baked at 60 ℃ for 15 min, then treated with Xylene Substitute (Citrisolv) (Fisher chemical cat. x3p-1GAL) then rehydrated and stained with Hematoxylin, followed with washes with running water. An incubation with Acid Alcohol (Sigma, cat. A3429) was performed, followed with washes with running water. The samples were then stained with eosin (Thermo Scientific) then dehydrated with ethanol followed by Xylene Substitute (Citrisolv). Samples were mounted with Permount (Fisher Scientific, cat. SP15-500). Slides were submitted for pathologic evaluation. Pathologist was blinded to the sample.
The mouse weight was recorded for analysis and lungs were collected for H&E staining. Pulmonary metastasis was evaluated by visually counting the number of metastatic nodules, maximum diameter per metastatic nodule and cross-section area per metastatic nodule in the entire mouse lung section for each mouse, using Nikon NIS-Elements software (Nikon Corporation Instruments, Tokyo, Japan).
The mouse weight was recorded for analysis and lungs were collected for H&E staining. Pulmonary metastasis was evaluated by visually counting the number of metastatic nodules, maximum diameter per metastatic nodule and cross-section area per metastatic nodule in the entire mouse lung section for each mouse, using Nikon NIS-Elements software (Nikon Corporation Instruments, Tokyo, Japan).
7
Histopathological Lung Tissue Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The degree of airway vascularity and extent of inflammation within the airways was assessed by histology. 5µm sections of lung specimens were stained with Harris haematoxylin (Sigma-Aldrich), washed using running tap water, then placed in acid alcohol (Sigma-Aldrich) to differentiate. Sections were then washed in running tap water and blued in Scott's tap water (Sigma-Aldrich). After rinsing in tap water, sections were stained with eosin (Sigma-Aldrich) then dehydrated through graded alcohols, cleared in xylene (MP Biomedicals Inc, Santa Ana, USA) and mounted using Faramount aqueous mounting medium (DakoCytomation, Glostrup, Denmark) and coverslipped. Sections were graded as per custom histopathology criteria designed by Horvat et al[10] (link).
8
Histological Analysis of Murine Osteosarcoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were sacri ced at four weeks (Experiment NO 1 and 2 in vivo) after secondary injection and the lung tissue were dissected. Osteosarcoma specimen of patients were stored at -80℃ after resection. The tissue was xed in 10% formalin at room temperature overnight and consequentially dehydrated in 70% ethanol. Samples were mounted in para n blocks and sectioned at 5 mm of thickness. Slides were baked at 60℃ for 15 minutes, then treated with Xylene Substitute (Citrisolv) (Fisher chemical cat. x3p-1GAL) then rehydrated and stained with Hematoxylin, followed with washes with running water. An incubation with Acid Alcohol (Sigma, cat. A3429) was performed, followed with washes with running water. The samples were then stained with eosin (Thermo Scienti c) then dehydrated with ethanol followed by Xylene Substitute (Citrisolv). Samples were mounted with Permount (Fisher Scienti c, cat. SP15-500). Slides were submitted for pathologic evaluation. Pathologist was blinded to the sample.
The mouse weight was recored for analysis and lungs were collected after H&E staining. Metastasis was measured by visually counting the number of metastatic nodules, maximum diameter per metastatic nodule and cross-section area per metastatic nodule in the entire mouse lung section for each mouse, using Nikon NIS-Elements software (Nikon Corporation Instruments, Tokyo, Japan).
The mouse weight was recored for analysis and lungs were collected after H&E staining. Metastasis was measured by visually counting the number of metastatic nodules, maximum diameter per metastatic nodule and cross-section area per metastatic nodule in the entire mouse lung section for each mouse, using Nikon NIS-Elements software (Nikon Corporation Instruments, Tokyo, Japan).
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!