Sperm concentration and mobility were assessed using a sperm counting chamber (Sperm Meter; Sperm Processor, Aurangabad, India) and light microscopy. Eosin/nigrosine staining was used to assess sperm morphology. Briefly, the epididymal sperm were washed in PBS, then 30 μl washed sperm were mixed with 60 μl eosin stain (Merck, Darmstadt, Germany) for 3 minutes. In the next step, 90 μl of nigrosine (Merck) was added to this mixture. For each sample, two smears were prepared, and 200 sperm were counted under an optical microscope. Abnormalities in the head, neck, and tail of sperm were determined, and a percentage of abnormal sperm morphology was reported for each sample [25 (link)].
Eosin stain
Manufactured by Merck Group
Sourced in United States, Germany
0.1% eosin stain is a laboratory staining solution used for microscopic analysis. It contains a concentration of 0.1% of the dye eosin, which is commonly used in histology and cytology to stain biological samples. The primary function of this stain is to provide contrast and enhance the visualization of cellular structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Hematoxylin, by Merck Group (2 mentions)
Formalin, by Merck Group (2 mentions)
Nigrosine, by Merck Group (1 mentions)
Lc ms grade solvents, by Merck Group (1 mentions)
Rhodamine b, by Merck Group (1 mentions)
Pluronic f 127, by Merck Group (1 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
Xylazine, by Merck Group (1 mentions)
Alginate powder, by Merck Group (1 mentions)
Hematoxylin stain, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Paraffin, by Merck Group (1 mentions)
Xylene, by Merck Group (1 mentions)
Malathion, by Merck Group (1 mentions)
Roswell park memorial institute (rpmi), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
16 protocols using eosin stain
1
Epididymal Sperm Morphology Assessment
2
Sorafenib Tosylate Formulation and Characterization
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Sorafenib tosylate (ST) was generously provided by Dr. Krishna Bhavanasi at Natco Pharma Ltd. (Kothur, Telangana, India). Monoolein (MO) (RYLO MG 19 Pharma) was sourced from Danisco Ingredients (Grinsted, Denmark). Pluronic F-127, Rhodamine B, and Streptozotocin (CAS ID 18883-66-4) were purchased from Sigma-Aldrich (Bangalore, India). Liquid chromatography–mass spectroscopy (LCMS)-grade solvents, hematoxylin, and eosin stain were obtained from Merck (Bangalore, India). Antibodies were procured from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and Abcam (Cambridge, MA, USA). All other chemicals utilized were of analytical reagent grade.
3
Sperm Viability Assessment via Eosin-Nigrosin Staining
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Sperm viability was evaluated by eosin-nigrosin staining according to a WHO protocol (25 ). In brief, 40 µL of eosin stain (1% in distilled water, Merck, Germany) was mixed to 20 µL sperm suspension. After 30 seconds, we added 60 µL of nigrosin stain (10% in distilled water, Merck, Germany). One drop of the mixture was placed on a microscope slide to generate a thin smear and examined under a light microscope at ×1000 magnification. In this method, viable spermatozoa remained colorless while nonviable spermatozoa stained red.
4
Histological Tissue Preparation Protocol
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The following chemicals including ethanol (Merck Company, Germany), paraffin, eosin stain, hematoxylin stain, xylene (Merck Company, Germany), formalin (Merck Company, Germany), acetic acid (Merck Company, Germany), ketamine HCl, xylazine, alginate powder, NaCl (Merck Company, Germany), distilled water, and HCl (Merck Company, Germany) were purchased.
5
Acetylcholinesterase Activity Assay
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Malathion (98% purity) (fyfanon 50 EC 500 g/l), acetylthiocholine iodide, 5.5’-dithiobis-(2-nitrobenzoic acid) (DTNB), Triton X-100, eosin stain, and RPMI (Roswell Park Memorial Institute), were purchased from SIGMA and Invitrogen.
6
Eosin Staining of Protoscoleces
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This examination was according the permeability of eosin stain (Sigma-Aldrich, St. Louis, MO, USA) solution (0.1%) in the protoscoleces and also the movement of flame cell. Dead protoscoleces are seen in red; nevertheless, the protoscoleces that are live do not absorb any color and will be seen colorless and typical activity of muscular and flame cell.
7
Colloidal Silver Nanoparticle Synthesis
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Colloidal Ag-NPs was obtained from Behban Shimi Pharmacy, Tehran, Iran, and were stored at room temperature (25°C) until testing. Amphotericin B, Eosin stain and Tween 20 were prepared from Sigma-Aldrich, (St. Louis, MO, USA). All the other chemicals and solvents used were of the highest purity commercially available.
8
Scolicidal Effect of N. sativa Extracts
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To determine the scolicidal activity of various extracts of N. sativa upon the protoscoleces of hydatid cysts, various concentrations of the extracts were used for 10, 20, 30 and 60 min. Initially, 0.5 ml of the protoscoleces (2× 103/ml) solution was placed in test tubes. In the next step, 0.5 ml of various concentrations of the extracts was added to each test tube, separately. The contents of the tubes were slowly mixed and then incubated at 37°C for 10, 20, 30 and 60 min. At the end of each incubation time, the upper phase was carefully removed so as not to interrupt the protoscoleces. Fifty µl of 0.1% eosin stain (Sigma-Aldrich, St Louis, MO, USA) was then added to the remaining settled protoscoleces and mixed gently. The upper portion of the solution was discarded after 10 min of incubation. The remaining pellet of protoscoleces was then smeared on a glass slide, covered with a cover glass and examined under a light microscope. The percentages of dead protoscoleces were determined by counting 200 protoscoleces. Furthermore, normal saline and 20% hypertonic saline were used as negative and positive control, respectively.
9
Evaluating Antiparasitic Effect of Zedoaria Essential Oil
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C. zedoaria essential oil with different concentrations (75, 150, and 300 µL/mL) (0.2 ml) were added to each test tube containing 0.2 ml of protoscoleces and then kept for 5, 10, 20 and 30 min at 37°C. After the incubation time, the supernatant was removed and 50 µL of 0.1% eosin stain (Sigma-Aldrich, St. Louis, MO, USA) was added to the protoscoleces. After 10 min of incubation, the protoscoleces were smeared on a glass slide, covered with a coverslip, and tested under a light microscope. The results were reported in the percentage of dead and live protoscoleces with the counting of 300 protoscoleces [14 (link)].
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C. zedoaria essential oil with different concentrations (75, 150, and 300 µL/mL) (0.2 ml) were added to each test tube containing 0.2 ml of protoscoleces and then kept for 5, 10, 20 and 30 min at 37°C. After the incubation time, the supernatant was removed and 50 µL of 0.1% eosin stain (Sigma-Aldrich, St. Louis, MO, USA) was added to the protoscoleces. After 10 min of incubation, the protoscoleces were smeared on a glass slide, covered with a coverslip, and tested under a light microscope. The results were reported in the percentage of dead and live protoscoleces with the counting of 300 protoscoleces [14 (link)].
10
Evaluating Zinc Nanoparticle Efficacy Against Protoscoleces
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After adding 0.5 ml of the protoscoleces solution to the laboratory tubes, 0.5 ml of ZnNPs at the concentrations of 50, 100, and 200 μg/ml alone and combined with albendazole (50 μg/ml) was put to the tubes and were keep warm for 5, 10, 20, 30, and 60 min. Once the incubation time was completed, 50 μl of 0.1% eosin stain (Sigma-Aldrich, St. Louis, MO, USA) was added to the remaining protoscoleces and smears were obtained on a glass slide, protected with a cover glass, and examined by a light microscope. The mortality rate of protoscoleces was recorded by counting 100 perotoscoleces; whereas normal saline and Ag nitrate were considered as negative and positive control [24 (link)].
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