Mice were anesthetized with CO2/O2 on gd 13.5 or 18.5, respectively. A blood sample was collected by retro bulbar puncture and subsequently mice were sacrificed by cervical dislocation. The harvested uterus-draining lymph node was kept in PBS on ice. The fetuses and corresponding placentas were isolated from the amniotic membranes. After assessment of fetal weight, fetuses were fixed with Bouin’s solution. The empty uterus was stored in HBSS on ice. Placentas were either stored at -20°C in RNAlater (Ambion by Life Technologies GmbH) for subsequent gene expression analysis or embedded in biopsy cassettes and stored in 4% Formaldehyde solution (36.5-38%, Sigma-Aldrich, St, Louis, US) for 24 h before transfer into 1% Formaldehyde solution for long-term storage and histological staining.
Formaldehyde solution
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, China
Formaldehyde solution is a versatile laboratory reagent used for various applications. It is a clear, colorless liquid that contains a concentrated solution of formaldehyde in water. The primary function of this product is to act as a fixative, preservative, and disinfectant in various laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (15 mentions)
Triton x 100, by Merck Group (14 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (14 mentions)
Sodium hydroxide (naoh), by Merck Group (11 mentions)
Hoechst 33342, by Thermo Fisher Scientific (11 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (11 mentions)
Rhodamine phalloidin, by Thermo Fisher Scientific (10 mentions)
Trypsin edta, by Thermo Fisher Scientific (9 mentions)
Resorcinol, by Merck Group (8 mentions)
Hydrochloric acid (hcl), by Merck Group (8 mentions)
Crystal violet solution, by Merck Group (7 mentions)
Methanol, by Merck Group (7 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (7 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (7 mentions)
Ethanol, by Merck Group (6 mentions)
Formic acid, by Thermo Fisher Scientific (6 mentions)
Lsm 510, by Zeiss (6 mentions)
Axio observer z1, by Zeiss (6 mentions)
Potassium chloride (kcl), by Merck Group (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (5 mentions)
245 protocols using formaldehyde solution
1
Maternal Immunology and Fetal Development
2
Maternal Physiology and Fetal Growth
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On gd 13.5 and 18.5, mice were anesthetized by carbon dioxide ventilation, a blood sample was collected by retro bulbar puncture and subsequently mice were sacrificed by cervical dislocation. The abdomen was opened and the uterus-draining lymph node was harvested and kept in PBS on ice. The uterus was removed and stored in HBSS on ice immediately after the fetuses were isolated from the amniotic membranes to determine fetal weight. Placentas were either stored at −20°C in RNAlater (Ambion by Life Technologies GmbH) or embedded in biopsy cassettes and stored in 4% Formaldehyde solution (36.5–38%, Sigma-Aldrich, St, Louis, US) for 24 h before transfer into 1% Formaldehyde solution for long-term storage. Maternal ovaries were also preserved in RNAlater.
3
Lipid Formulation for Photodynamic Therapy
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1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was obtained from Avanti Polar Lipids (Alabaster, AL). 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol (DSPE-PEG, average PEG molecular mass of 2,000 amu), ZnPC (97 % purity), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), pyridine, and sulforhodamine B (SRB) were acquired from Sigma-Aldrich (St. Louis, MO). Acetic acid (glacial), ethidium bromide, ethylenediaminetetraacetic acid (EDTA), formaldehyde solution (36.5–38 % in water), sodium chloride, and tris(hydroxymethyl)aminomethane (Tris) were obtained from Merck KGaA (Darmstadt, Germany). Agarose was purchased from Gibco-BRL (Paisley, UK) and ethanol was from J.T. Baker (Deventer, the Netherlands).
All lipids were dissolved in chloroform and ZnPC was dissolved in pyridine at a 178-μM concentration. All dissolved lipids were stored under a nitrogen atmosphere at −20 °C.
All lipids were dissolved in chloroform and ZnPC was dissolved in pyridine at a 178-μM concentration. All dissolved lipids were stored under a nitrogen atmosphere at −20 °C.
4
Evaluating Drug Synergy in Pancreatic Cancer
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Indicated cells were cultured and seeded into 96-well plates at a density of 300–2,000 cells per well, depending on growth rate. Twenty-four hours later, drugs were added at the indicated concentrations using the HP D300 Digital Dispenser (HP). After 72 h, medium and drugs were refreshed. The total duration of the experiment was 6 days (two treatments) for KCP_K2101, KCP_P0012, MiaPaCa-2 and Panc10.05, and 10 days (three treatments) for Panc1, ASPC1 and YAPC. Cells were fixed with 4% PFA diluted in PBS (37% Formaldehyde solution, Merck) and stained with 2% crystal violet solution (HT90132-1 L, Sigma-Aldrich). Drug synergy was calculated using CompuSyn software (version 1.0), which is based on the median-effect principle and the combination index–isobologram theorem.64 (link) CompuSyn software generates combination index values, where combination index 0–0.75 indicates synergy, 0.75–1.25 indicates an additive effect and CI > 1.25 indicates antagonism.65 (link) Following the instructions of the software, drug combinations at non-constant ratios were used to calculate the combination index in our study.
5
Histological Evaluation of Tissue Samples
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At the end of each experimental period, the animals were anesthetized and killed by administrating high doses of anesthetics. Shaving and incision were performed and the tubes and surrounding tissues were removed with a safety margin of 1 cm. The specimens were kept in 10% formaldehyde solution (Merck & Co, Darmstadt, Germany) for 2 weeks, until they were histologically processed.
The polyethylene tubes were removed from the specimens, which were then set in prepared paraffin blocks. Sections 5 µm thick were cut by a microtome and stained with hematoxylin and eosin and three sections from each specimen were selected.
The polyethylene tubes were removed from the specimens, which were then set in prepared paraffin blocks. Sections 5 µm thick were cut by a microtome and stained with hematoxylin and eosin and three sections from each specimen were selected.
6
Viral Plaque Assay for Quantification
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In order to verify the absence of plaque in the βPL-inactivated samples and quantify the viral stock used in animal challenge, 24-well plates with pre-formed monolayers of Vero CCL-81 cells were inoculated with 4-fold serial dilutions of the samples (100 to 10−6, in duplicate). After 1 h of adsorption (37 °C, 5% CO2), the inoculum was removed, and 1 mL of semi-solid medium, produced from medium 199 (supplemented with 5% FBS, 40 µg/mL of gentamicin sulfate, and 1 µg/mL of amphotericin B (Gibco, Billings, MA, USA)), and 1.5% medium viscosity carboxymethylcellulose sodium salt (Sigma-Aldrich, Saint Luis, MO, USA) were added to each well, and the plates were incubated for 72 h at 37 °C, 5% CO2. Cell monolayers were fixed by adding 1 mL of 1.25% formaldehyde solution (Merck, Darmstadt, HE, Germany) to each well and incubating the plates for at least 90 min (final concentration of 0.625% formaldehyde). Following the fixation step, the plates were washed and stained with 0.4% crystal violet solution (Sigma-Aldrich, Saint Luis, MO, USA). Plaque counts were performed, and the viral titer was calculated in Log10 PFU/mL.
7
T Cell Activation Assay Protocol
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Culture medium (CM) was prepared using RPMI 1640 supplemented with 100 U/mL of penicillin, 100 μg/mg streptomycin (both from Sigma, Steinheim, Germany), and 2 mM L-glutamine (Biochrom, Berlin, Germany). Concanavalin A (ConA) (Sigma) was diluted in CM to a concentration of 600 μg/mL and stored at −70°C. Antibodies were purchased from Becton Dickinson (BD, Heidelberg, Germany). Phosphate buffered saline (PBS) was made by dissolving 7.013 g NaCl, 0.2 g KCl, 1.513 g Na2HPO4, and 0.2 g KH2PO4 (all purchased from Roth) in 1 liter distilled water and by adjusting the pH to 7.4. Red blood cell (RBC) lysing solution was purchased from Becton Dickinson and diluted 1 : 10 in distilled water before use. Washing buffer was obtained from BD Biosciences (San Diego, USA). Formaldehyde solution and absolute methanol was purchased from Merck (Darmstadt, Germany).
8
Cytoskeletal Effects of Dental Sealers
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To assess variations in the morphology, structure, and organization of the F-actin cytoskeleton of hPDLSCs under exposure to the different sealer eluates, a qualitative description of immunofluorescence images of phalloidin-stained cells was performed. To do so, hPDLSCs were seeded onto glass coverslips, left to adhere, and cultured in DMEM (control) or in DMEM treated with 1:1, 1:2, or 1:4 of BrF, AHPbcs, or AHP for 72 h at 37ºC. Then, the following was performed: 1) cells were rinsed twice using pre-warmed foetal bovine serum at 37ºC; 2) cells were fixed in 4% formaldehyde solution (Merck Millipore, Darmstadt, Germany) for 10 min; 3) cells were made permeable with 0.25% Triton X-100 solution (Sigma-Aldrich) for 5 min; and 4) cell cytoskeleton and nuclei were stained with AlexaFluor™594-conjugated phalloidin (Invitrogen) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (ThermoFisher Scientific, Waltham, MA, United States), respectively. Lastly, immunofluorescence images were obtained and observed under a confocal microscope (Leica TCS SP2; Leica, Wetzlar, Germany). Each experimental condition and visualization were performed in triplicate.
9
Thymoquinone Effects on Collagen Production
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Thymoquinone was purchased from Sigma-Aldrich (CAS number: 490-91-5); hydroxy-proline kit was purchased from Kiazist (Kiazist Co., Hamedan, Iran). Formaldehyde solution was purchased from Merck; cryotube was purchased from Bio Plas Inc., xylazine and ketamine were purchased from Nasr (Iran), and rabbits were purchased from the animal center of Shiraz University of Medical Science.
10
Histopathological Evaluation of Lung Tissue
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Lung specimens were fixed in formaldehyde solution 4%, buffered, pH 6.9 (Merck, Germany). Lung tissue samples were dehydrated and were embedded in paraffin (Merck, Germany) and sectioned into 5 μm thickness slices with 25 μm intervals. A usual hematoxylin and eosin (H&E) staining was performed for histological assessment. Five sections from each corpse were used for evaluations. Two histopathologists, blinded to the groups, evaluated the sections. The sections were visible under a light microscope (LABOMED, Labo America, Inc. USA). A digital camera (LABOMED, USA) was used to take histophotographs. Total histopathological scoring was done according to the following parameters: hemorrhage, thickened wall, inflammatory cells infiltration, and fibrotic changes with a score of None = 0, Mild = 1, Moderate = 2, and Severe = 3 (Gibson-Corley et al. 2013 (link)).
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