Picric acid, a highly explosice chemical compound, is a key reagent in various research fields.
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After myocardial infarction and four weeks of reperfusion, hearts were either paraffin embedded or the raw material was snap frozen at -40°C in isopentane. In paraffin sections (4 µm) the wax was dissolved by an organic solvent and the tissue slices rehydrated before picrosirius red stain was applied. Cryosections (8 µm) from raw snap frozen tissue were fixed in 4% paraformaldehyde (PFA) in 0.1M sodium phosphate buffer (PB) pH 7.4 or Zambonis fixative (0.1 M PB, 4% (w/v) PFA, 15% (v/v) picric acid) for 10 min. Washing steps were performed in PBS, PBS/0.1% Saponin or in PBS/0.2% Tween 20 according to the further requirements. Picrosirius red (SR) staining was performed according to the protocols of Junqueira et al.3 (link) and Sweat et al.5 (link)Wheat germ agglutinin (WGA) labeling was routinely used in combination with the secondary antibody in immunohistochemical preparations. Lectin from triticum vulgaris FITC conjugate (# L4895, Sigma-Aldrich, St. Louis, MO, USA) was diluted 1:100 (10 µg/mL) in the required buffer. Incubation time was one hour protected from light. After three washing steps, the sections were coverslipped with a water-soluble antifading mounting medium. Collagen I staining was performed using an anti-collagen I antibody (ab34710, Abcam, Cambridge, UK) diluted 1:100 in the required buffer. As secondary antibody anti-Rabbit-Cy3 (111-165-144, Jackson Dianova, Hamburg, Germany) was used in a concentration of 2.5 µg/mL. Slides were analyzed with fluorescence microscope Keyence BZ 9000 (Keyence, NeuIsenburg, Germany). All shown images were taken with 4x objective using the merge function, as not otherwise specified.
Emde B., Heinen A., Gödecke A, & Bottermann K. (2014). Wheat Germ Agglutinin Staining as a Suitable Method for Detection and Quantification of Fibrosis in Cardiac Tissue after Myocardial Infarction. European Journal of Histochemistry : EJH, 58(4), 2448.
Collagen fibers were detected in colonic tissues as previously reported [23 (link)]. Briefly, colonic samples were incubated in 0.04% Fast Green for 15 min, washed with distilled water and then incubated in 0.1% Fast Green and 0.04% Sirius Red in saturated picric acid for 30 min. Then, they were dehydrated and mounted with DPX Mounting. Collagen fibers appeared red, while the non-collagen proteins were green. Quantitative estimations of histochemical stainings were carried out independently by two blind investigators (C.S. and C.I.). Each investigator analyzed all tissue specimens under study. The respective values were then averaged and plotted in graphs in accordance with previously described criteria [23 (link)]. Briefly, for each animal, 5 randomly selected microscopic fields from 3 non-adjacent sections, which were selected every 18 sections in order to ensure the evaluation of colonic samples with an average thickness of 160 μm, were captured by a Leica DMRB microscope equipped with the digital camera DFC480. All images, which were captured with 100x or 400x objective for studying the whole wall or tunica muscularis, respectively, were quantitatively estimated for collagen fibers and cellular non-collagen proteins in double Sirius Red/Fast Green staining, within the respective colonic areas. To detect the specific threshold of different colors (e.g., pink/red for collagen fibers and green for non-collagen proteins), a square was applied upon the color of interest and recorded by Image Analysis System ‘L.A.S. software v.4’. Positive tissue areas were automatically estimated on the basis of the total pixel number and intensity. The whole wall and tunica muscularis areas were manually circumscribed and automatically calculated. Data were expressed as percentage of Σ of positive-stained area / Σ of tissue area examined of whole wall or tunica muscularis in three colonic sections (5 fields/each) for each animal.
Segnani C., Ippolito C., Antonioli L., Pellegrini C., Blandizzi C., Dolfi A, & Bernardini N. (2015). Histochemical Detection of Collagen Fibers by Sirius Red/Fast Green Is More Sensitive than van Gieson or Sirius Red Alone in Normal and Inflamed Rat Colon. PLoS ONE, 10(12), e0144630.
In vivo studies utilized a transgenic mouse engineered to express the HIV-1 Tat1-86 protein in astrocytes in a doxycyline-inducible manner. As previously described (Bruce-Keller et al. 2008 ), mice expressing the Tat gene under control of a tet responsive element (TRE) in the pTREX vector (Clonetech, Mountain View, CA) were crossed with mice engineered to express the glial fibrillary acidic protein (GFAP) promoter driving the reverse tetracycline transactivator (RTTA). The inducible Tat transgenic mice (Tat+ mice) used in these studies express both GFAP-RTTA and TRE-Tat genes, while the control mice (Tat- mice) express only the GFAP-RTTA gene. Individual animals were genotyped using RNA isolated from ear clip samples, by standard PCR procedures previously described (Bruce-Keller et al. 2008 ). Tat-induction was obtained by replacing standard chow with a formulation containing doxycycline (DOX) at 6 mg/g (Harlan, Indianapolis, IN). Some control groups were also fed DOX-containing chow to control for non-specific actions of DOX intake. Morphine was delivered by 25 mg, timed-release pellets implanted subcutaneously in the subscapular region (NIDA Drug Supply System) under aseptic conditions using isoflurane anaesthesia. Sham pellets were similarly implanted as controls. The 25 mg morphine pellets reportedly deplete at a rate of 5 mg/day, yet steady state levels of morphine, measured in ng/ml plasma or ng/gm brain tissue reflect lower availability, probably due to the balance of delivery and metabolism (see, e.g., (Feng et al. 2006 (link))). Both C3HeB and C57Bl mice reportedly tolerate delivery from 75 mg/5 day morphine pellets (Peart and Gross 2004 (link); Rahim et al. 2003 (link)). Less than 5% of the mice in our studies died, and there was no relationship between morphine-induced toxicity and either genotype or DOX administration. In experiments lasting for a total of 7 days, opioid and control pellets were replaced after 5 days. Naltrexone was administered via an Alzet mini-pump (1007D) implanted in the same region. 600 mg/ml naltrexone (NIDA Drug Supply System) was prepared in 50% DMSO, a standard vehicle for mini-pump delivery (Arnot et al. 1996 (link)). 100 μl total volume was loaded per mini-pump, delivering 0.5 μl/hr for up to 7 days. Pumps containing 50% DMSO and sterile saline were used as naltrexone controls. For studies where mice were exposed to both opioids and Tat, mice were given the DOX feed starting on the night before opioid treatment in order for blood levels of DOX to stabilize. For histological studies, mice were deeply anaesthetized with halothane prior to perfusion with Zamboni's fixative (2% paraformaldehyde, pH 7.4, with 0.15% picric acid).
Hauser K.F., Hahn Y.K., Adjan V.V., Zou S., Buch S.K., Nath A., Bruce-Keller A.J, & Knapp P.E. (2009). HIV-1 Tat and Morphine Have Interactive Effects on Oligodendrocyte Survival and Morphology. Glia, 57(2), 194-206.
Blood samples were collected from all cows all at one time. The samples were collected in the morning, before the feed distribution, by venipuncture from the jugular vein, using 10-mL Li-heparin treated tubes (Vacuette, containing 18 IU of Li-heparin mL−1, Kremsmünster, Austria). Samples were immediately cooled in an ice-water bath after collection. The blood was centrifuged (3500 × g for 16 min at 4 °C) and the plasma obtained was separated into two aliquots: the first fraction was immediately used to collect the infrared spectra; the second one was stocked at −20 °C until metabolites analysis and the results were used as calibration values. Plasma metabolites used as calibrating values were analyzed at 37 °C by an automated clinical analyzer (ILAB 600, Instrumentation Laboratory, Lexington, MA), using the methodology showed in Table 1. Commercial kits were used to measure glucose, total cholesterol, urea, calcium, inorganic phosphorus, magnesium, total protein, albumin, total bilirubin, and creatinine (Instrumentation Laboratory SpA, Werfen, Monza, Milan, Italy), NEFA and zinc (Wako, Chemicals GmbH, Neuss, Germany), and β-OH-butyric acid (BHBA, kit Ranbut, Randox Laboratories Limited, Crumlin, County Antrim, United Kingdom Randox, UK). A Trinder end point [Glucose oxidase (GOD)/Peroxidase (POD)] was used to measure glucose. Total cholesterol (cholesterol and cholesterol esters) was also measured using Trinder end point [Cholesterol oxidase (CHOD)/Peroxidase (POD)], after a hydrolysis of cholesterol esters to free cholesterol. Urea was measured with end point method using the couple urease glutamate dehydrogenase (GLDH) enzyme system. Colorimetric methodology based on the reaction of calcium with o-cresolphthalein complexone was used to measure Ca with end point method. Inorganic phosphorus was measured with end point UV method, based on the reaction between phosphate ions in an acidic medium with ammonium molybdate to form a phosphomolybdate complex. The magnesium determination was based on the reaction of magnesium with Xylidyl Blue (as chelator) at alkaline pH, which yields a purple colored complex. Total protein were measured with the modified biuret methodology, based on the reaction of peptide bonds with Cu++ ions in alkaline solution to form a colored complex. Albumin was measured with an end point colorimetric method, based on the binding between albumin with green bromocresol resulting in a spectral change of the dye from yellow to green. Total bilirubin was determined with an end point analysis using modified Jendrassik-Grof method, based on the reaction between total bilirubin with diazotized sulfanilic acid in presence of lithium dodecylsulfate to form azobilirubin. Creatinine was analyzed with an end point colorimetric method, based on the reaction of creatinine with picric acid under alkaline conditions. NEFA were determined with an Trinder end point [Acyl coenzyme A oxidase (ACOD)/Peroxidase (POD)] assay, after the acylation of coenzyme A by NEFA contained in the sample. BHBA was measured with a kinetic UV method, based on the oxidation od D-3 hydroxybutyrate to acetoacetate by 3-Hydroxybutyrate dehydrogenase. Electrolytes, Na, K, and Cl, were measured using a potentiometric system, with specific electrodes. Kinetic analysis was adopted to determine the activity of enzymes: alkaline phosphatase (AP, EC 3.1.3.1), aspartate aminotransferase (AST, EC 2.6.1.1), γ-glutamyltransferase (GGT, EC 2.3.2.2) using Instrumentation Laboratory kits (Instrumentation Laboratory SpA, Werfen, Monza, Milan, Italy). Ceruloplasmin was measured using the method described by Sunderman and Nomoto [16 (link)], adapted to ILAB 600 condition. The method is based on measurement of p-phenylenediamine dihydrochloride oxidation by the oxidase activity of ceruloplasmin. Finally, haptoglobin was measured using the method described by Skinner et al. [17 (link)] and Owen et al. [18 (link)] and adapted to ILAB 600 condition. The method is based on peroxidase activity of methaemoglobin-haptoglobin complex measured by the rate of oxidation of guaiacol (hydrogen donor) in presence of hydrogen peroxide (oxidizing substrate).
Methodologies used to measure the plasma parameters with reference methods
Parameter
Methodology
Wavelength (nm)
CVa
Glucose
Endpoint
510
1.50
Total cholesterol
Endpoint
510
2.10
NEFAb
Endpoint
546
1.50
BHBAc
Endpoint
340
5.25
Urea
Endpoint
340
1.20
Creatinine
Endpoint
510
5.40
Ca
Endpoint
570
1.40
Inorganig P
Endpoint
340
2.00
Mg
Rate
340
1.40
Na
ISE deviceg
0.90
K
ISE deviceg
1.30
Cl
ISE deviceg
1.50
Zn
Endpoint
546
Ceruloplasmin
Endpoint
546
3.48
Total protein
Endpoint
546
1.20
Albumin
Endpoint
600
1.80
Total bilirubin
Endpoint
546
6.70
Haptoglobin
Endpoint
450
13.54
ASTd
Rate
340
2.10
GGTe
Rate
405
3.72
APf
Rate
405
1.70
aCalculared on the results obtained between runs according to the National Committee for Clinical Laboratory Standards (Document EP3-T: Guidelines for Manufacturers for Establishing
Performance Claims for Clinical Chemistry Methods, Replication Experiment
Evaluation”, Villanova, PA, 1982.)
bNon esterified fatty acids;
cβ-OH-butyric acid;
dAspartate amino transferase
eγ-glutamyl transferase
fAlkaline phosphatase
gIon selective electrodes
Calamari L., Ferrari A., Minuti A, & Trevisi E. (2016). Assessment of the main plasma parameters included in a metabolic profile of dairy cow based on Fourier Transform mid-infrared spectroscopy: preliminary results. BMC Veterinary Research, 12, 4.
Hearts were excised, washed with saline solution, and placed in 10% formalin. Hearts were then cut transversely close to the apex to visualize the left ventricle and right ventricle. Several sections of heart (5 μm thick) were prepared and stained with hematoxylin and eosin and a saturated solution of picric acid containing 1% Sirius red for collagen deposition (18 (link)). The sections were then visualized by light microscopy and photographed, and the collagen content of the sections was measured by using the computer-assisted morphometry (Image-Pro Plus Version 6.0). For each sample, all available fields (>30 fields) were measured, including the septum and the right and the left ventricle (all fields were analyzed with a ×40 objective lens). For cardiomyocyte cross-sectional area, sections were stained for membranes with fluorescein isothiocyanate–conjugated wheat germ agglutinin (WGA; Invitrogen) and for nuclei with DAPI (19 (link)). A single cardiomyocyte was measured with an image quantitative digital analysis system (NIH Image version 1.6). The outline of 200 cardiomyocytes was traced in each section. Tissue sections (5 μm) were stained with antibodies against tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-α1, and GRP78, respectively. Detection was carried out by using the EnVision+ system and diaminobenzidine (USCNLIFE, China) as described previously (20 (link)).
Li J., Zhu H., Shen E., Wan L., Arnold J.M, & Peng T. (2010). Deficiency of Rac1 Blocks NADPH Oxidase Activation, Inhibits Endoplasmic Reticulum Stress, and Reduces Myocardial Remodeling in a Mouse Model of Type 1 Diabetes. Diabetes, 59(8), 2033-2042.
HBFP staining was used to evaluate myocardial infarction in rats after CME. The 4-micron ventricular myocardial paraffin sections were dewaxed, followed by hematoxylin staining, alkaline fuchsin staining, and differentiating in picric acid-acetone solution. Finally, the sections were dehydrated and sealed. The results were observed through an optical microscope (ZEISS, Axio Imager.A2, Germany, 200 times magnification). Ischemic myocardium and erythrocytes were stained with red, and the normal myocardium was stained with yellow or brownish yellow, then the nuclei were stained with blue black. The measurements of infarct size were performed using the Image J (NIH, MD, USA) software.
Li H., Yang H., Qin Z., Wang Q, & Li L. (2024). Colchicine ameliorates myocardial injury induced by coronary microembolization through suppressing pyroptosis via the AMPK/SIRT1/NLRP3 signaling pathway. BMC Cardiovascular Disorders, 24, 23.
Procedure: In a glass tube, 100µL of the extract at a dose of 1000µg/ml was introduced. Then, 1000µL of 13% picric acid and 1000µL of 4% NaOH were added respectively. The tubes were boiled in a water bath for 10 minutes. In the white tube, the extract was replaced by distilled water. After the tubes were cooled, the optical density was read at 570nm against the blank and the extract concentration was obtained from the glucose calibration curve and expressed as glucose equivalent per mg of dry plant extract.
Hervé B., Estella T.F., Legrand N.B.N., Ngameni B., & Charles F.N. (2024). Phytochemical Screening and In vitro Antimicrobial Activity of Four Cameroonian Medicinal Plant Extracts against Bacterial and Fungal Strains Involved in Skin Lesion Infections. Journal of Advances in Medical and Pharmaceutical Sciences, 26(3), 9-24.
Sirius Red staining was performed on 20 µm-thick cryosections of EDL muscles. Sections were incubated overnight at RT in a Bouin Solution (saturated picric acid, 37% formaldehyde, 5% acetic acid). After washing in running water, sections were incubated in a solution of 0.1% Direct Red 80 in saturated picric acid for 1 h at RT. After washing with 2% acetic acid, sections were dehydrated in sequence with 40-50% ethanol, 70% ethanol, 96% ethanol, 100% ethanol, 1:1 ethanol-xylene (100%), 100% xylene. Slides were mounted with Eukitt (Sigma).
Di Marco G., Gherardi G., De Mario A., Piazza I., Baraldo M., Mattarei A., Blaauw B., Rizzuto R., De Stefani D, & Mammucari C. (2024). The mitochondrial ATP-dependent potassium channel (mitoKATP) controls skeletal muscle structure and function. Cell Death & Disease, 15(1), 58.
The picric acid test is used to detect reducing sugars. First, 1 mL of the saturated picric acid was added to 1 mL of the test honey samples, followed by 0.5 mL of 10% sodium carbonate (10%) solution. Then, the test tube was heated in a boiling water bath and the absorbance was read at 490 nm [82 (link)].
where A sample = Sample absorbance, and A standard = Standard absorbance.
Hameed O.M., Shaker O.M., Ben Slima A, & Makni M. (2024). Biochemical Profiling and Physicochemical and Biological Valorization of Iraqi Honey: A Comprehensive Analysis. Molecules, 29(3), 671.
Test for HCN production was carried out by the method as described by [42 ]. A sterilized nutrient broth amended with glycine (4.4 g/l), already poured into the sterile test tubes, was inoculated by 24 h old bacterial culture. The filter paper (Whatman No. 1) was cut into strips and immersed in 0.5% picric acid. These strips were inserted between the plug and the inner wall of the test tube so that it hung above the inoculated broth. Inoculated tubes were sealed with parafilm tape to hold the gaseous metabolic produced by the antagonistic bacteria and allow for a chemical reaction with picric acid on the top. After incubation for a week at 28 °C, the colour of the filter paper was observed. The positive result was indicated by the color shift of a filter paper strip immersed in picric acid from yellow to red.
Bhargavi G., Arya M., Jambhulkar P.P., Singh A., Rout A.K., Behera B.K., Chaturvedi S.K, & Singh A.K. (2024). Evaluation of biocontrol efficacy of rhizosphere dwelling bacteria for management of Fusarium wilt and Botrytis gray mold of chickpea. BMC Genomic Data, 25, 7.
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Direct Red 80 is a synthetic dye commonly used in laboratory settings. It is a water-soluble, red-colored azo dye that is utilized in various applications, such as staining and coloring procedures. The core function of Direct Red 80 is to provide a consistent and reliable coloring agent for specific laboratory techniques and processes. Further details on its intended use are not available.
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Picric acid is a yellowish crystalline compound that is used as a laboratory reagent. It is a powerful oxidizing agent and has applications in various chemical processes and analyses.
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Sirius Red is a lab equipment product manufactured by the Merck Group. It is a dye used for the detection and quantification of collagen in biological samples. Sirius Red binds specifically to collagen fibers, allowing for their visualization and analysis.
The Ultracut-S microtome is a laboratory instrument designed for the precise cutting of ultra-thin sections, typically used in electron microscopy sample preparation. It features precise control and high-quality sample sectioning capabilities.
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Fast Green is a laboratory staining dye used in various scientific applications. It is a synthetic, water-soluble dye that provides a green coloration. The core function of Fast Green is to stain and visualize specific components or structures within biological samples during microscopy and other analytical procedures.
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Permount is a mounting medium used in microscopy to permanently mount specimens on glass slides. It is a solvent-based, xylene-containing solution that dries to form a clear, resinous film, securing the specimen in place and providing optical clarity for microscopic examination.
Sirius Red F3B is a synthetic dye used in the laboratory for the staining of collagen fibers. It binds specifically to collagen and produces a bright red color, allowing for the visualization and analysis of collagen content in various tissue samples.
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Sodium hydroxide is a chemical compound with the formula NaOH. It is a white, odorless, crystalline solid that is highly soluble in water and is a strong base. It is commonly used in various laboratory applications as a reagent.
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Paraformaldehyde is a white, crystalline solid compound that is a polymer of formaldehyde. It is commonly used as a fixative in histology and microscopy applications to preserve biological samples.
Saturated picric acid is a chemical compound used in various laboratory applications. It serves as a sensitive reagent for the detection and identification of certain organic compounds. The core function of saturated picric acid is to provide a colorimetric, or visible, reaction when mixed with specific substances, allowing for their identification and analysis.
Picric acid is a highly explosive chemical compound, which poses significant safety risks. Proper handling, storage, and disposal procedures are critical when working with this substance. Additionally, the availability and accessibility of Picric acid can be limited in certain regions due to regulations, making it challenging to obtain for research purposes.
Yes, there are several variations or types of Picric acid, each with its own unique properties and applications. For example, Potassium Picrate is a salt form of Picric acid that is less sensitive to shock and friction, making it potentially safer to handle in some cases. Researchers should familiarize themselves with the specific properties and requirements of the Picric acid variant they are using.
Picric acid is a versatile chemical compound with a wide range of applications in research. It is commonly used as a reagent in analytical chemistry, organic synthesis, and various biochemical assays. Picric acid is also employed in the production of certain explosives and pyrotechnics, though these applications are often highly regulated.
PubComapre.ai allows you to screen protocol literature more efficiently and leverage AI to pinpoint critical insights. The platform can help researchers identify the most effective protocols related to Picric acid for their specific research goals. PubCompare.ai's AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy, while also addressing the unique challenges associated with working with this explosive compound.
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More about "Picric acid"
Picric acid, also known as 2,4,6-Trinitrophenol or Lyddite, is a highly explosive chemical compound that has important applications in various research fields.
This yellow crystalline solid is a powerful oxidizing agent and a popular reagent in diverse scientific disciplines, including analytical chemistry, histology, and explosives research.
In analytical chemistry, picric acid is used as a colorimetric reagent for the detection and quantification of certain metal ions, such as iron and copper.
It is also employed in the preparation of dyes and stains, like Direct Red 80, Sirius Red, and Fast Green, which are commonly used in histological and microscopy techniques.
In the field of histology, picric acid is a key component in fixation and staining procedures.
When combined with formaldehyde, as in the Bouin's solution, it forms a powerful fixative that helps preserve tissue structure and morphology.
The picric acid-based Sirius Red stain is widely used to identify collagen fibers in tissues, making it a valuable tool for researchers studying the extracellular matrix and connective tissue disorders.
The versatility of picric acid extends to the realm of explosives research, where it serves as a model compound for the study of explosive behaviors and the development of safer handling protocols.
The Ultracut-S microtome, a specialized instrument used for sectioning materials, including energetic compounds like picric acid, plays a crucial role in this field of study.
Despite its utility, picric acid must be handled with great caution due to its highly reactive and explosive nature.
Appropriate safety measures, such as the use of Permount mounting medium, are essential when working with this chemical.
Proper disposal methods, such as treatment with sodium hydroxide to form the less reactive sodium picrate, are also crucial to ensure the safe handling of picric acid.
In summary, picric acid is a versatile and important chemical compound that finds applications in various research fields, from analytical chemistry to histology and explosives research.
Researchers can leverage the power of AI-driven platforms like PubCompare.ai to streamline their picric acid-related workflows, identify the best protocols and products, and enhance the reproducibility and accuracy of their studies.
