For antigen unmasking, the slides were boiled in 10 mM sodium citrate buffer (Gibco). The slides were treated with 3% hydrogen peroxide (Biosesang Corp., Sungnam, Korea) for 15 min to reduce endogenous peroxidase and with normal goat serum to minimize non-specific binding. To detect the proliferative cells, slides were treated by anti-mouse ki67 IgG and biotinylated anti-rabbit antibody (Cell Signaling Technology, Danvers, MA, USA). The stained tissues by avidin/biotinylated enzyme complex kit (Vector Laboratories, Inc., Burlingame, CA, UK) were developed using the 3,3-diaminobenzidine substrate (Sigma). The slides of each group were evaluated by randomly selecting 5 photographs per mice. Digital images were obtained using the Leica Application Suite Microscope Software (Leica Microsystems Inc.). The used magnification was × 200.
Application suite microscope software
Manufactured by Leica
Sourced in Germany
Leica Application Suite Microscope Software is a comprehensive digital imaging solution that provides an integrated platform for acquiring, processing, analyzing, and managing microscopic images. The software offers a user-friendly interface, advanced imaging tools, and a range of features to support various microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
Mayer s paracarmine, by Merck Group (2 mentions)
Sodium citrate buffer, by Thermo Fisher Scientific (1 mentions)
Avidin biotinylated enzyme complex kit, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine substrate, by Merck Group (1 mentions)
Dm 1000 led microscope, by Leica (1 mentions)
Stereoscan model s 2469n scanning electron microscope, by Hitachi (1 mentions)
Neutralized formalin, by Merck Group (1 mentions)
Stereoscan model s 2469n, by Hitachi (1 mentions)
6 protocols using application suite microscope software
1
Antigen Retrieval and Proliferation Assay
2
Morphological Analysis of Parasite Specimens
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The specimens were stained with Mayer’s paracarmine (Merck, Darmstadt, Germany), dehydrated in an ethanol series, cleared in methyl salicylate and mounted in Canada balsam for morphological analysis. Specimens were then examined using a compound microscope equipped with a bright field (Leica DM 1000 LED microscope, Leica, Wetzlar, Germany). Measurements were taken using Leica Application Suite microscope software (Leica Microsystems GmbH, Wetzlar, Germany) and are given in micrometres as range followed by mean in parentheses. Some specimens from each locality were dehydrated in ethanol series, critical-point dried, sputter coated with gold, and examined with a Hitachi Stereoscan Model S-2469N scanning electron microscope operating at 15 kV. Adults and metacercariae were deposited in the Colección Nacional de Helmintos (CNHE) of the Instituto de Biología, Universidad Nacional Autónoma de México (UNAM), México City (CNHE. No. 12063–12066).
3
Histological Analysis of Hair Regrowth
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Dorsal skin from all mice was fixed in 10% neutralized formalin (Sigma) for 18 h. Fixed skins were dehydrated with ethanol and xylene. 4 μm cut paraffin-embedded sections were prepared with rotary microtome. Hematoxylin and eosin (H&E) solution (YD Diagnostics Corp., Yongin, Korea) was applied to the slides to monitor the hair regrowth. The slides of each group were evaluated by randomly selecting 5 photographs per mice. Digital images were obtained using the Leica Application Suite Microscope Software (Leica Microsystems Inc., IL, USA). The used magnification was × 100.
4
Staining and Microscopic Analysis of Digeneans
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Digeneans preserved in 100% ethanol were stained with Mayer's paracarmine (Merck, Darmstadt, Germany), dehydrated in ethanol series, cleared in methyl salicylate and mounted in Canada balsam for morphological analysis. Specimens were examined using a compound microscope equipped with a bright field Leica DM 1000 light emitting diode microscope (Leica, Wetzlar, Germany). Measurements were taken using Leica Application Suite microscope software (Leica Microsystems GmbH, Wetzlar, Germany) and are given in micrometres and presented with the range followed by the mean in parentheses. Some specimens were dehydrated with an ethanol series, critical point dried, sputter coated with gold and examined with a Hitachi Stereoscan Model S-2469N scanning electron microscope operating at 15 kV. Voucher specimens from the present study were deposited in the Colección Nacional de Helmintos (CNHE) from Instituto de Biología, Universidad Nacional Autónoma de México (UNAM), Mexico City.
5
Plaque Assay for Adenovirus Quantification
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A549 and LN18 monolayers seeded in 6-well plates were infected with serial dilutions of rAds. Two hours post-infection, the viral inoculum (400 µl) was removed and cells were covered with 3 ml of a mix of DMEM/5% FBS/1% agarose. Two days later, DMEM/5% FBS overlay was added. To evaluate the plaque size, monolayers were stained six (A549) or ten (LN18) days post-infection by incubating with 0.5 mg/ml thiazolyl blue tetrazolium bromide (MTT) at 37 ºC for 4 h. The plaques were photographed at 50 × with the Leica DM3000 microscope and quantified with a Leica Application Suite microscope software.
6
Plaque Assay for Recombinant Adenovirus
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A549 monolayers seeded in 6-well plates were infected with serial dilutions of rAds. Two hours post-infection, the viral inoculum was removed, and the cells were covered with 2–3 mL of a mix of DMEM/5% FBS/1% agarose. Later, DMEM/5% FBS overlay was added. For evaluation of the plaque size, monolayers were stained by incubating with a 1/10th volume of thiazolyl blue tetrazolium bromide (MTT, 0.5 mg/mL) at 37°C for 4 h. The plaques were photographed at 50× with a Leica DM3000 microscope and quantified with Leica Application Suite microscope software.
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