Oocyte diameter was measured using an ocular micrometer calibrated with a stage micrometer fitted in a light microscope (Labex, India). For each prawn, the diameter of at least 30 oocytes was measured and the mean oocyte diameter was calculated. The stage of oocyte development was characterized based on the maximum number of oocytes confined to a particular stage of development. Photomicrographs of various stages of oocyte development were taken using Leica 2500 microscope (Germany).
2500 microscope
Manufactured by Leica
Sourced in Germany
The Leica 2500 microscope is a high-performance optical instrument designed for scientific and industrial applications. It features a stable, vibration-resistant stand, a precise focusing mechanism, and a range of objective lenses to accommodate various magnification requirements. The microscope provides clear, detailed images for observation and analysis purposes.
Automatically generated - may contain errors
8 protocols using 2500 microscope
1
Oocyte Morphometry and Development
2
Histological Analysis of Ovary Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Triplicate histological analyses were done by sacrificing three animals from each group. For this, ovary was dissected out carefully. The tissue samples were fixed in Bouin's fixative for 24 h and washed with distilled water. The samples were dehydrated with different graded alcohol series and processed by routine procedure. Sections of 6–8 μm thickness were made and stained with haematoxyline and eosin. The stained sections were mounted using Dibutyl phthalate xylene (DPX) and photomicrographs of varying magnifications were taken using Leica 2500 microscope.
3
Wound Healing and Migration Assays for PASMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For wound healing assay, when the density of PASMCs reached 90–100%, the cells were starved for 24 h, and then scratched with a 100 μL pipette tip. After incubating with PDGF-BB with/without SIL or XMA for 48 h, the number of cell migration was calculated under a phase Leica 2500 microscope. For the transwell migration chamber assay, PDGF-BB with/without SIL or XMA were added to the lower chamber when PASMCs (5 × 104 cells in 100 μL serum-free medium) were placed in the upper chamber. Forty-eight hours later, the unmigrated cells were gently removed from the top, and the cells migrated to the submembrane were fixed with 4% paraformaldehyde, stained with 5% crystal violet, and counted under the Leica 2500 microscope.
4
Quantifying Renal Histopathology in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collected kidneys were immediately fixed in 4% polyformaldehyde (pH 7.4) for 24 h, embedded in paraffin, and then sliced into 4–6 μm sections. After haematoxylin and eosin (HE) staining, 10 glomeruli per rat were randomly chosen for calculating the glomerular surface area using Image-Pro Plus software 6.0. To evaluate the mesangial matrix area (%), periodic acid-Schiff (PAS) staining was employed according to the protocol [16 (link)]. Ten glomeruli from each rat were analysed on a digital microscope screen grid containing 540 points in Adobe Photoshop Element 6.0. The percentage of relative mesangial matrix area was calculated by (number of grid points in the mesangial area)/(total number of points in the glomerulus). To evaluate the degree of renal fibrosis, the Masson trichrome stain (MTS) was used as described previously [25 (link)]. The sample sections were photographed randomly with a Leica 2500 microscope (Leica Microsystems, Wetzlar, Germany), and the percent of fibrotic areas was quantified using Image-Pro Plus software 6.0.
5
Histopathological Analysis of Rat Lung and RV
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collected rat lung tissues and RV tissues were immediately fixed in 4% polyformaldehyde (pH 7.4) for 24 h, embedded in paraffin, and then sliced into 4 μm sections. Then lung sections were stained with hematoxylin and eosin (H&E), toluidine blue (TB), picrosirius red (PSR) staining, Masson trichrome staining (MTS), and primary antibody against CD68 (Abcam, MA, USA) according to the protocols described previously [17 (link)]. Pulmonary arterial wall thickness (PAWT) was determined according to the ratio of (external diameter − internal diameter) to external diameter based on H&E staining. The right ventricular hypertrophy was determined by the cross section area (CSA) of cardiomyocytes using H&E staining. Graphs were observed under a Leica 2500 microscope (Leica Microsystems, Wetzlar, Germany).
6
Histological Analysis of Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lung tissue samples were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned at 5 μm intervals. Hematoxylin-eosin (H&E), Elastica van Gieson (EVG), and Masson trichrome staining (MTS) were performed using standard procedures. The sections were then examined and photographed with a Leica 2500 microscope (Leica Microsystems, Wetzlar, Germany). The pulmonary arterial wall thickness was calculated as follows: medial wall thickness (%) = (external diameter–internal diameter)/external diameter × 100. For quantitative analysis, 20 randomly selected vessels with an external diameter of 25–100 μm from each rat were analyzed.
7
In situ Visualization of H2O2 and O2⦁- Accumulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H2O2 accumulation was monitored in situ with 3,3′-diaminobenzidine tetrachloride (DAB; 1997) . Six-day-old seedlings were immersed in DAB (1 mg ml -1 ) in PBS buffer (10 mM; pH 7.0) plus 0.05% (v/v) tween-20 and placed overnight at room temperature in the dark. O2 •- was detected based on nitroblue tetrazolium (NBT)-reducing activity. Seedlings were covered in an NBT solution (0.5mg ml -1 ) in 0.1M PBS buffer (pH 7.4) plus 0.05% triton-X 100 and incubated in the light for 10 minutes. DAB and NBT reactions were stopped by replacing staining solution with fixative / clearing solution of ethanol / acetic acid / glycerol (3:1:1) for 24 hours, mounted on glass slides with a coverslip and visualized with a Leica 2500 microscope (www.leica-microsystems.com).
8
Quantifying Lung Injury and Permeability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse lung tissue was fixed with 4% polyformaldehyde and embedded in paraffin, sliced into 5-µm sections, stained with hematoxylin and eosin (H&E) and subjected to immunohistochemical staining with a primary antibody against Ly6G (Abcam). The lung injury score was calculated using H&E staining according to a formula described in an American Thoracic Society report [21] . Permeability of the alveolar capillary membranes were assessed using an Evans blue dye (EB) extravasation technique [22] . Briefly, mice were given an intraperitoneal injection with EB dye (50 ml/kg) 2 h before sacrifice. At the end of exposure, the pulmonary circulation was flushed. The EB dye extracted from lung tissue were then determined at 620 and 750 nm using a microplate reader (Thermo Scientific, CA, USA). Cellular apoptosis in lung tissue from different groups was determined using a TUNEL assay kit (In Situ Cell Death Detection Kit, POD, Roche Company, Germany) described previously [23] . The TUNEL-positive cells were stained green, and the number of TUNEL-positive cells in eight different random fields from each section was counted using a Leica 2500 microscope (Leica Microsystems, Wetzlar, Germany) at ×200 magnification.
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!