Fluorescein-isothiocyanate dextran of 4 kDa (FD4) and 59–77 kDa (FD70), Alcian blue 8Gx, and Hank's Balanced Salt Solution (HBSS) were purchased from Sigma-Aldrich (Søborg, Denmark). Sodium decanoate > 99% (C10) and MES anhydrous BioChemica (MES) were acquired from Tokyo Chemical Industry Co. (Tokyo, Japan) and PanReac AppliChem (Darmstadt, Germany), respectively. Sucrose was bought from WVR (Søborg, Denmark), and Ringer tablets, periodic acid solution, and Schiff's reagent were obtained from Merck KGaA (Darmstadt, Germany). SPHERO™ Fluorescent Nile Red Particles with a diameter of 0.1–0.3 µm were purchased from Nordic BioSite (Copenhagen, Denmark). Mannitol D-[1-14C] (57.2 mCi/mmol) and Ultima Gold scintillation fluid were purchased from Perkin Elmer (Ballerup, Denmark). Metoprolol-[3H] (27.6 Ci/mmol) was obtained from Vitrax Radiochemicals (Placentia, CA, USA). Alexa Fluor 555-conjugated oligonucleotides targeting bacterial 16S RNA (sequence: 5′-GCTGCCTCCCGTAGGAGT-3′) were synthesized by Eurofins Genomics (Ebersberg, Germany). Primers for intestinal gene expression were designed based on published sequences of target genes in pigs using the NCBI online primer design tool (Primer-BLAST: http://www.ncbi.nlm.nih.gov/tools/primer-blast/ ). Freshly prepared ultrapure water (18.2 MΩ × cm) purified by a PURELAB flex 4 (ELGA High Wycombe, UK) was used if not otherwise stated.
Periodic acid solution
Manufactured by Merck Group
Sourced in United States
Periodic acid solution is a laboratory reagent used in various analytical and synthetic chemistry procedures. It is a clear, colorless liquid containing periodic acid (HIO4) dissolved in water. The primary function of this solution is to serve as an oxidizing agent in organic synthesis and analytical techniques. Specific applications and usage instructions should be obtained from the manufacturer or relevant scientific literature.
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Lab products found in correlation
Schiff s reagent, by Merck Group (16 mentions)
Schiff reagent, by Merck Group (9 mentions)
Alcian blue 8gx, by Merck Group (2 mentions)
Roti histokitt, by Carl Roth (2 mentions)
Gills number 3 haematoxylin, by Merck Group (2 mentions)
Hematoxylin, by Merck Group (2 mentions)
D 1 14c mannitol, by PerkinElmer (1 mentions)
Ringer tablets, by Merck Group (1 mentions)
Hank s balanced salt solution, by Merck Group (1 mentions)
Ultima gold scintillation fluid, by PerkinElmer (1 mentions)
Dm5500 widefield microscope, by Leica (1 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Ix70 microscope, by Olympus (1 mentions)
Ix2 ucb microscope, by Olympus (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Oncostatin m, by Merck Group (1 mentions)
Luxol fast blue solution, by Merck Group (1 mentions)
Genegenius bioimaging system, by Syngene (1 mentions)
34 protocols using periodic acid solution
1
Fluorescent Tracer Protocols for Intestinal Permeability
2
Preservation of Intestinal Mucus for Histological Analysis
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The water-free methacarn (methanol-Carnoy’s) fixation was used to preserve the intestinal mucus layer during histological preparation.67 (link) Intestinal tissues with fecal pellets were fixed over night with methacarn at room temperature. Tissues were washed in methanol for 30 min, two times in ethanol for 15 min, once in ethanol/xylene (1:1) for 15 min and two times in xylene for 15 min prior to embedding in paraffin.
For periodic-acid-Schiff (PAS) histology, 3–5 µm thin tissue sections were dewaxed and stained for 10 min with periodic acid solution (0.5%, w/v; Merck), 20–30 min in Schiff’s reagent (Merck) and 1 min in Mayer’s Hämalaun (Carl Roth). Between each step, sections were rinsed in running water.
For FISH analysis, 3–5 µm thin tissue sections were dewaxed and then treated with 50 µl 4% lysozyme solution (45 min, 37°C) to demask nucleic acids. After washing, 50 µl hybridization solution was added and incubated for 3 hours at 50°C. Slides were washed several times at 37°C and were dried at RT before mounted with ProLong™ Gold Antifade Mountant with DAPI (Thermo Fisher Scientific) following manufacturer’s instructions. The samples were documented at a Leica DM 5500 wide field microscope (Leica) and analyzed with ImageJ (National Institutes of Health).
For periodic-acid-Schiff (PAS) histology, 3–5 µm thin tissue sections were dewaxed and stained for 10 min with periodic acid solution (0.5%, w/v; Merck), 20–30 min in Schiff’s reagent (Merck) and 1 min in Mayer’s Hämalaun (Carl Roth). Between each step, sections were rinsed in running water.
For FISH analysis, 3–5 µm thin tissue sections were dewaxed and then treated with 50 µl 4% lysozyme solution (45 min, 37°C) to demask nucleic acids. After washing, 50 µl hybridization solution was added and incubated for 3 hours at 50°C. Slides were washed several times at 37°C and were dried at RT before mounted with ProLong™ Gold Antifade Mountant with DAPI (Thermo Fisher Scientific) following manufacturer’s instructions. The samples were documented at a Leica DM 5500 wide field microscope (Leica) and analyzed with ImageJ (National Institutes of Health).
3
Glycogen Assessment in Placental Stem Cells
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Example 8
This example describes exemplary methods for assessing glycogen production by hepatocytes obtained from differentiated placental stem cells. Following depolymerization, cells are transferred to tissue culture treated 24 well plates (Falcon, BD Biosciences) and fixed with 10% formalin-ethanol fixative solution for 15 minutes at room temperature, with subsequent washes with PBS. Fixed cells are exposed to 0.25 ml of Periodic Acid Solution (Sigma Aldrich) per well for 5 minutes at room temperature. Glycols are oxidized to aldehydes in this process. After washing cells with PBS to remove the PAS, 1 ml of Schiff's reagent is added per well and cells exposed for 15 minutes at room temperature. Schiff's reagent, a mixture of pararosaniline and sodium metabisulfite, reacts to release a pararosaniline product that stains the glycol-containing cellular elements. A third PBS wash to remove the reagent is followed by image acquisition with an Olympus IX70 microscope and Olympus digital camera.
4
Periodic Acid-Schiff Staining Protocol
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Paraffin sections (5 μm) were deparaffinized and hydrated to deionized water. Next, slides were immersed in periodic acid solution (Sigma-Aldrich, St. Louis, MI, USA) for 5 min at room temperature. Then, the slides were rinsed several times in distilled water and immersed in Schiff’s Reagent (Sigma-Aldrich, St. Louis, MI, USA) for 15 min. At the end of the incubation, slides were washed in running tap water for 5 min, dehydrated, mounted with ROTI®Histokitt (Carl Roth GmbH, Karlsruhe, Germany), and imaged on the Olympus IX2-UCB microscope.
5
Periodic Acid-Schiff Staining of Colon Sections
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1% Periodic acid solution (Sigma-Aldrich) was used to incubate deparaffinised colonic sections for 10 min. And then Schiff reagent (Sigma-Aldrich) was used for incubation for 40 min. The PAS-stained sections were counterstained with Hematoxylin for 2–5 min. Extensive PBS solution was used to rinse well between each step.
6
Tissue Histological and Immunofluorescence Analysis
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Tissues were fixed in 10% buffered formalin and embedded in paraffin. Five μm paraffin sections were deparaffinized and rehydrated. For Periodic acid–Schiff staining, rehydrated tissue sections were treated with 0.5% periodic acid solution (Sigma-Aldrich, 3951), stained with Schiff's reagent (Sigma-Aldrich, 3952016), dehydrated and mounted. Immunofluorescence analysis was performed as described (19 (link),21 ) with anti-GR (C-terminal, IGBMC, #3249) and anti-Pax7 (DSHB, AB_528428) antibodies. Mouse or rabbit IgGs were used as controls.
7
Histological Tissue Analysis via H&E and PAS Staining
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The muscle tissues were fixed in 10% neutral buffered formalin, embedded in paraffin, and sectioned. The sections were stained with hematoxylin and eosin (H&E) or periodic acid-Schiff (PAS) in accordance with the following procedure: briefly, slides were deparaffinized by incubation in xylene, hydrated by washes in a graded series of ethanol solutions (100%, 95%, 80%, and 70%), washed in distilled water, and then stained with H&E. For PAS staining, the tissue sections were incubated with periodic acid solution (Sigma-Aldrich, St. Louis, MO, USA) for 20 min followed by Schiff solution for 20 min.
8
Hepatic Differentiation of Mesenchymal Stem Cells
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The materials used in the study included sodium alginate (medium viscosity; mannuronic acid content 50% from Nova Matrix Co.); trypsin/EDTA; fibroblast growth factor 4 (FGF4); hepatocyte growth factor (HGF); phosphate-buffered saline (PBS); glucagon; insulin-transferrin-selenium (ITS) supplement (100X); Dulbecco's modified Eagle's medium (DMEM); oncostatin M; dexamethasone (Dex); dimethyl sulfoxide (DMSO); trichostatin A (TSA) from Sigma; fetal bovine serum (FBS) from Gibco; penicillin/streptomycin; L-ascorbic acid 2-phosphate; enzyme-linked immunosorbent assay (ELISA) kit (Pars Azmun); urea assay kit (Pars Azmun); periodic acid solution; and Schiff (Sigma).
The study protocol was approved by the Ethics Committee of Ahvaz Jundishapur University of Medical Sciences (AJUMS), and all procedures were performed according to the ethical committee approval (Ethical code: IR.AJUMS.REC1395.267).
The study protocol was approved by the Ethics Committee of Ahvaz Jundishapur University of Medical Sciences (AJUMS), and all procedures were performed according to the ethical committee approval (Ethical code: IR.AJUMS.REC1395.267).
9
Histopathological Analysis of Canine Globoid Cell Leukodystrophy
10
Luxol Fast Blue Staining of Mouse Brain
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Sections of mouse brains (6 μm) were deparaffinized in xylene and hydrated in descending ethanol concentrations. Sections were then incubated in 0.1% Luxol Fast Blue Solution (Sigma) at 56 0C for 16–18 h, allowed to cool at RT, rinsed for 5 min in 95% ethanol in distilled water to remove excess blue stain. Sections were subsequently placed in 0.1% lithium carbonate aqueous solution for 2 min, dipped several times in 70% ethanol in distilled water in order to stop the reaction. Sections were then transferred to a chamber containing 0.8% periodic acid solution (Sigma) for 10 min and finally stained in Schiff’s reagent for 20 min, followed by 3 washes in a buffer containing 0.5% HCL (vol/vol) and 0.2% potassium disulfite (w/vol) for 2 min each. Sections were mounted with coverslips and then observed using light microscopy.
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