All femoral muscle tissue samples were fixed with 4% paraformaldehyde and embedded in paraffin wax. After slicing the fixed femoral muscle tissue samples and staining them with Hematoxilin–Eosin (HE), their physiological characteristics were observed and photographed under a microscope for subsequent analysis. The details of this three‐step process (embedding, slicing, and staining with HE) were as follows. First, the fixed tissue samples were washed with sterile water for 30 min and dehydrated in a tissue embedding plastic basket using 75% ethanol for 6 h, 85% ethanol for 10 h, 95% ethanol for 4 h, anhydrous ethanol I for 2 h, anhydrous ethanol II for 2 h, through transparent (Xylene I 20 min, xylene II 15 min) and soaked with paraffin wax for 3 h, resulting in tissue blocks embedded in paraffin wax. Second, a Leica RM2235 slicer was used to cut the femoral muscle tissue samples into 5 μm‐thick slices. After the tissue slices were flattened in warm water, they were placed on slides and baked at 60℃ for at least 2 h. Finally, the tissue slices were dewaxed with xylene and washed with water for 20 min. After staining the dewaxed tissue slices with hematoxylin for 30 min and with eosin for 5 min, the stained slices were sealed with a resin adhesive.
Rm2235 slicer
Manufactured by Leica
The RM2235 slicer is a microtome designed for sectioning tissue samples. It features a manual operation and can produce sections of varying thickness. The instrument is suitable for use in various laboratory applications requiring tissue sectioning.
Automatically generated - may contain errors
5 protocols using rm2235 slicer
1
Histological Analysis of Femoral Muscles
2
Detailed Immunofluorescence Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TriPure RNA Isolation Reagent, a Transcriptor First Strand cDNA Synthesis Kit, and SYBR Premix EX Taq II (2 ) were purchased from Roche. The RM2235
slicer and the Feather Microtome Blade 35 slicer were purchased from Leica.
The BX51 positive fluorescence microscope was purchased from Olympus.
The xylene, anhydrous ethanol, 4 % ethanol polyformaldehyde, 1 % phosphate-buffered saline (PBS)
solution, citric acid buffer, and anti-fluorescence attenuation tablets used were
all of analytical purity. The primary antibody used was rabbit anti-EPHA4 antibody
(BA2275-1). The secondary antibodies used were goat anti-rabbit IgG/Cy3 conjugated antibody, rabbit anti-Ephrin A3 primary antibody (bs-9758R, Bolson), and anti-rabbit
IgG secondary antibody (whole molecule)-FITC (Sigma).
slicer and the Feather Microtome Blade 35 slicer were purchased from Leica.
The BX51 positive fluorescence microscope was purchased from Olympus.
The xylene, anhydrous ethanol, 4 % ethanol polyformaldehyde, 1 % phosphate-buffered saline (PBS)
solution, citric acid buffer, and anti-fluorescence attenuation tablets used were
all of analytical purity. The primary antibody used was rabbit anti-EPHA4 antibody
(BA2275-1). The secondary antibodies used were goat anti-rabbit IgG/Cy3 conjugated antibody, rabbit anti-Ephrin A3 primary antibody (bs-9758R, Bolson), and anti-rabbit
IgG secondary antibody (whole molecule)-FITC (Sigma).
3
Ileum Histomorphometry and Goblet Cell Density
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ileum tissue samples were fixed in 4% paraformaldehyde, then embedding in paraffin. Each ileum sample was cut into 5 μm thick slide by Leica RM2235 slicer, each sample had 5 slides and each slide had 3 sections, then stained with the periodic acid Schiff method (PAS staining). Images were taken under the microscope. Ten well-oriented villi-crypts units of each section (Image ProPlus 6.0) were measured and villi–crypt ratio (VCR) was calculated. Image ProPlus 6.0 was used to count the goblet cells and measure the tissue areas, then calculate the goblet cell density (number of cells/µm2), and goblet cells density were obtained from 3 sections by each small intestinal segment.
4
Soybean Seedling Histological Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The top buds of soybean seedlings in the same growth period were selected, dissected with an anatomical needle and fixed in formaldehyde-acetic acid-ethanol (FAA) for 24 h. The treatments were as follows: 50%, 70%, and 85% alcohol solutions were used to dehydrate each tissue for 30 min, samples were then placed in 95% alcohol containing 0.5% eosin for 3 h, and anhydrous alcohol for 1 h (2 times); the alcohol solutions of 25%, 50%, and 75% were used for xylene transparentizing for 30 min, and 100% xylene for 30 min (3 times). After treatment in 50% xylene + 50% paraffin, samples were embedded within pure wax in a 60 °C oven. The samples were sliced using a Leica RM 2235 slicer, displayed and baked in an oven. Then, slices were dewaxed with xylene, alcohol, and distilled water, mordanted with 4% iron alum for 30 min, dyed with 0.5% hematoxylin, and finally treated with 2% alum for color separation. After washing and returning to blue, slices were dehydrated with alcohol solution. After being redyed in 95% alcohol containing 1% eosin, and after being sealed, the slices were observed under an Olympus BX 51 optical microscope. Photoshop was used to prepare the pictures [45 (link)].
5
Kernel Development in Maize Mutants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immature wild-type and dek39 mutant kernels were collected from the same segregating ears of selfpollinated heterozygous plants at 12 DAP. The kernel was cut along the longitudinal axis into three equal parts. The central slice containing the embryo was fixed at 4°C in 4% paraformaldehyde overnight. The fixed material was dehydrated in a graded ethanol series (50, 70, 80, 95, and 100% ethanol) . After clearing with HistoChoice Clearing Agent (Sigma, H2779) and infiltrating with paraffin wax, the sample was embedded and sectioned at 8 mm with a Leica RM2235 slicer. The sections were stained with Hematoxylin and Eosin, and observed with Olympus SZ51 microscopy.
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!