MSCs at an initial density of 1 × 104 cells/cm2 were seeded onto different membranes for 14 and 21 d. At each time point, cells were fixed with 4% paraformaldehyde for 1 h and then stained with alizarin Red S solution for 45 min in the dark. Subsequently, the samples were washed with ddH2O until no more stains appeared in the washing solution. Then, the samples were imaged using a stereoscopic microscope (NIKON, Japan). The stained calcium nodules were subjected to 10% cetylpyridinium chloride for 30 min for quantitative analysis. The dissolved staining solution was extracted and transferred to a new 96-well plate, and the corresponding OD values were recorded at 540 nm. Furthermore, the mineralized nodules on different membranes after 21 d were further observed by SEM.
Stereoscopic microscope
Manufactured by Nikon
Sourced in Japan
The Nikon Stereoscopic Microscope is a high-performance optical instrument designed for detailed observation of specimens. It provides a three-dimensional, magnified view of the subject matter, enabling users to examine the depth, texture, and fine details with clarity. The stereoscopic design allows for binocular viewing, enhancing the depth perception and overall clarity of the observed sample.
Automatically generated - may contain errors
11 protocols using stereoscopic microscope
1
Quantitative Analysis of Mineralized Nodules
2
Preparation of DRG Neurons for Patch-Clamp
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Before DRG neuron preparation, glass coverslips (10 mm round, Assistant, Germany) were first cleaned by 75% (vol/vol) ethanol and air dried in a sterile culture hood. Cleaned coverslips were coated with poly-D-lysine (0.1 mg/ml, 90 μl per coverslip, O/N) and laminin (20 μg/ml, 5 μl per coverslip). Coated coverslips were sterilized again with UV light for 10 min before plating into the sterile 60 mm central well dish.
Seven days after SNI surgery or three days after microRNA agomir/antagomir microinjection, L4 and L5 DGRs were used for whole-cell patch clamp recording. After animals were euthanized with isoflurane, fresh DRGs ipsilateral and contralateral to SNI surgery site were quickly isolated and placed into cold Neurobasal Medium. The excess nerves and blood vessels were dissected under a stereoscopic microscope (Nikon, Japan) using micro scissors and forceps. The isolated DRGs were dissociated with 1 mg/ml collagenase type I and 2 mg/ml dispase in Neurobasal Medium for 30–35 min. DRG cell suspension was added to the dishes with pre-coated coverslips and placed into the incubator (37 °C, 5% CO2) for 2–4 h before recording.
Seven days after SNI surgery or three days after microRNA agomir/antagomir microinjection, L4 and L5 DGRs were used for whole-cell patch clamp recording. After animals were euthanized with isoflurane, fresh DRGs ipsilateral and contralateral to SNI surgery site were quickly isolated and placed into cold Neurobasal Medium. The excess nerves and blood vessels were dissected under a stereoscopic microscope (Nikon, Japan) using micro scissors and forceps. The isolated DRGs were dissociated with 1 mg/ml collagenase type I and 2 mg/ml dispase in Neurobasal Medium for 30–35 min. DRG cell suspension was added to the dishes with pre-coated coverslips and placed into the incubator (37 °C, 5% CO2) for 2–4 h before recording.
3
Analyzing Triploid Block in Arabidopsis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dry seeds from five siliques were collected and imaged under a stereoscopic microscope (Nikon), and the triploid block was quantified by counting the number of aborted seeds. The same set of seeds was then surface-sterilized using 50% bleach and ethanol, rinsed once with ethanol 96%, and air-dried. The seeds were sown on agar plates containing 0.5X MS medium, 1% sucrose, pH = 5.7, stratified for 2 days at 4°C, and transferred to growth chambers at 23°C, 70% humidity, 120 μE m−2 light with a 16-h light/8-h dark (long days) photoperiod germination rate was initially quantified on 4- to 5-days-old seedlings, then adjusted after 7 days if necessary to account for germination delays.
4
Quantifying MSC Colony Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the ability to produce colony-forming units, single-cell suspensions (1 × 103 cells) were seeded in 10 cm diameter culture dishes. After 14 days of culture, three MSCs were fixed in 4% paraformaldehyde and stained with 0.1% toluidine blue for 30 min. After being washed with phosphate-buffered solution (PBS), images were taken by stereoscopic microscope (Nikon, Japan).
5
Embryonic Mouse Tissue Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryonic and neonatal mice were sacrificed on ice. Photos of embryos were taken with a stereoscopic microscope (Nikon, Japan).
After the bodies had been fixed in 4% paraformaldehyde for over 24 h, they were dehydrated in a series of increasingly concentrated sucrose solutions. Then, frozen sections were generated according to a previously described protocol and stored at −20 °C.14 (link) These sections were brought to room temperature for the immunofluorescence analysis.
After the bodies had been fixed in 4% paraformaldehyde for over 24 h, they were dehydrated in a series of increasingly concentrated sucrose solutions. Then, frozen sections were generated according to a previously described protocol and stored at −20 °C.14 (link) These sections were brought to room temperature for the immunofluorescence analysis.
6
Osteogenic Potential of MSCs on Membranes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MSCs at an initial density of 1 × 104 cells/cm2 were seeded onto different membranes for 4 and 7 d. For ALP staining to be visualized, at each predetermined time point, cells were fixed with 4% paraformaldehyde for 1 h, and then stained following the protocol provided by 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium (BICP/NBP) Alkaline Phosphatase Color Development Kit (Beyotime, China). Finally, the stained membranes were captured using a stereoscopic microscope (NIKON, Japan). For the ALP activity assay, cells were subjected to lysis using 1% Triton X-100 in ice for 40 min. The supernatant was collected by centrifuging at 12,000 rpm for 10 min. The total protein content and ALP activity were measured using a Bicinchoninic acid assay kit (Beyotime, China) and an alkaline phosphatase assay kit (Nanjing Jiancheng Bioengineering Institution, China). The OD values for protein and ALP detection were recorded using a spectrophotometric microplate reader (Bio-Rad 680, USA) at 562 nm and 520 nm.
7
Investigating Afngg1's Effect on A. flavus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To investigate the effect of Afngg1 on the growth of A. flavus, 2 μL spore suspensions (106 spores/mL) of A. flavus control, ΔAfngg1 and ΔAfngg1-Com strains were separately inoculated in the centre of PDA plates and cultured at 30 °C for 5 days; then, the colony diameter was measured and conidial heads were counted under a stereoscopic microscope (Nikon, Shanghai, China). Next, 6 mL sterile water was added to each Petri dish to harvest the mycelium, and the number of conidia was calculated using a hemocytometer. To assess the hydrophobicity of A. flavus mycelia, 20 μL sterile water and 20 μL 3% Bromophenol Blue solution were added to the edges of colonies of A. flavus control, ΔAfngg1 and ΔAfngg1-Com strains, cells were cultured at 30 °C for 2 days, and analysed by a stereoscopic microscope. Quantitative analysis of sclerotia production was performed at 37 °C for 7 days. After 7 days, 75% ethanol was sprayed to wash the medium and expose the sclerotia, and five holes were made along the diameter to count the number of sclerotia. Additionally, scanning electron microscopy (SEM) was used to photograph the conidia microstructure of A. flavus control, ΔAfngg1 and ΔAfngg1-Com strains as described previously [58 (link)].
8
C. elegans Growth and Reproduction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
According to previous literature [17 (link)], the C. elegans was treated at 50 °C for 10 min after exposure to ethanol extract from PHS of different concentrations for 24 h, observed and photographed under stereoscopic microscope (Nikon, Japan), and measured the length of C. elegans by Image J software (Bethesda, MD, USA). Thirty C. elegans were measured in each concentration group.
Thirty C. elegans exposed to different concentrations of PHS ethanol extract for 24 h were selected into the normal NGM, one in each medium, and transferred the medium every 12 h until the end of the spawning period of C. elegans [18 (link)]. Record the number of all offspring that it hatches as an indicator of the number of offspring.
Thirty C. elegans exposed to different concentrations of PHS ethanol extract for 24 h were selected into the normal NGM, one in each medium, and transferred the medium every 12 h until the end of the spawning period of C. elegans [18 (link)]. Record the number of all offspring that it hatches as an indicator of the number of offspring.
9
Larval Development of Aquatic Species
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
From fertilized eggs, 50 eggs were sampled at random and put in a 500 mL glass beaker, and weak aeration was maintained. Breeding water temperature was maintained at 24.5~26.5℃ (average 25.0±0.05℃), and half of the breeding water was changed twice a day. For egg size, egg diameter was measured up to 0.01 mm using a phase-contrast microscope (LEICA, Germany), and eggs developing after fertilization were observed longitudinally using a stereoscopic microscope (NIKON, Japan). Larvae just after hatching were held in a rectangular glass tank (30×25×30 cm), and bred through recirculation using a sponge filter. During the period of breeding, the larvae and juveniles were fed with Artemia from just after hatching to 20 days, and then with the mixture of Daphnia pulex and formula feed (700 μm). For the morphological development of larva and juvenile, everyday from hatching, five individuals were sampled and anesthetized using anesthetic (MS-222, Ethyl 3-aminobenzoate methanesulfonate, Sigma Aldrich Co., St. Louis, USA), and the developmental characteristics of each part of the body were examined using a stereoscopic microscope and profile projector (NIKON, Japan). The stages of morphological development were divided according to Kendall (1984) .
10
Histochemical and Enzymatic Analysis of GUS Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histochemical assay of GUS and enzymatic activity assay were performed according to Jefferson et al. [52 (link)]. The plants were incubated overnight at 37 °C in 2 mM X-Gluc (5-bromo-4-chloro-3-indolyl b-D-glucuronide) with a phosphate buffer (pH 7.0) containing 10 mM ethylenediaminetetraacetic acid (EDTA), 0.5 mM potassium ferricyanide, 0.5 mM potassium ferrocyanide, and 0.1% v/v Triton X-100. By using 70% ethanol after X-Gluc staining, chlorophyll was completely removed from the plants, which were then photographed using a stereoscopic microscope (Nikon, Tokyo, Japan).
The procedure for the GUS fluorometric assay was performed according to Niu et al. [53 (link)]. GUS protein can react with the substrate 4-methylumbelliferyl-β-d-glucuronic acid (MUG, Sangon, Shanghai, China) to generate 4-MU (4-methylumbelliferone). GUS enzymatic activity is expressed as pmol 4-MU produced per milligram protein per minute. GUS activity was measured using a Varioskan Flash Spectral Scanning Multimode Reader (Thermo Fisher, Waltham, MA, USA) with 365 nm excitation and 455 nm emission.
The procedure for the GUS fluorometric assay was performed according to Niu et al. [53 (link)]. GUS protein can react with the substrate 4-methylumbelliferyl-β-d-glucuronic acid (MUG, Sangon, Shanghai, China) to generate 4-MU (4-methylumbelliferone). GUS enzymatic activity is expressed as pmol 4-MU produced per milligram protein per minute. GUS activity was measured using a Varioskan Flash Spectral Scanning Multimode Reader (Thermo Fisher, Waltham, MA, USA) with 365 nm excitation and 455 nm emission.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!