After fixation in 4% paraformaldehyde for 24 h, the ilea were soaked through a graded series of ethanol and xylene, embedded in paraffin, and sectioned at 5 μm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL). The sections were deparaffinized with xylene and rehydrated through graded dilutions of ethanol and stained with hematoxylin and eosin. The images of ilea were acquired using an Olympus simon-01 microscope (Olympus Optical Co., Ltd., Beijing, China). The values of villus height and crypt depth were measured 5 times from different villus and crypts per section from each broiler using the Image-Pro Plus software 6.0 (Media Cybernetics, Inc., Washington, DC). The average value of the 5 villus and crypts represent the value of each ileum sample.
Lecia rm2235 microtome
Manufactured by Leica
Sourced in United States
The Leica RM2235 is a rotary microtome designed for precise sectioning of paraffin-embedded tissue samples. It features a motorized operation with adjustable cutting speed and section thickness range from 0.5 to 60 micrometers. The Leica RM2235 is a versatile instrument suitable for a variety of histological and pathological applications.
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9 protocols using lecia rm2235 microtome
1
Histological Analysis of Broiler Ilea
2
Histological Analysis of Liver and Ovary
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After fixation in 4% paraformaldehyde for 24 h, the liver and ovary were soaked through a graded series of ethanol and xylene, embedded in paraffin, and sectioned at 4 μm with a Lecia RM2235 microtome (Leica Biosystems, Buffalo Grove, IL). The sections were deparaffinized with xylene and rehydrated through a graded dilutions of ethanol, and stained with hematoxylin and eosin (HE ). The images of the liver and ovary were acquired using an Olympus simon-01 microscope (Olympus Optical, Beijing, China).
3
Histological Analysis of Broiler Jejunum
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After fixation in 4% paraformaldehyde for 24 h, the jejunal sections were soaked through a graded series of ethanol and xylene and embedded in paraffin. The jejuna were sectioned at 5 μm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL). The sections were deparaffinized with xylene and rehydrated through a graded dilution of ethanol. Hematoxylin and eosin staining and Alcian blue-periodic acid Schiff staining were performed, respectively. The images of jejuna were acquired using an Olympus simon-01 microscope (Olympus Optical Co., Ltd., Beijing, China). The values of villus height (VH ), crypt depth (CD ), and the number of jejunal goblet cells were measured 5 times from different villus and crypts per section from each broiler using the Image-Pro Plus software 6.0 (Media Cybernetics, Inc., Rockville, MD).
4
Histological Analysis of Intestinal Morphology
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After fixation in 4% paraformaldehyde for 24 h, the jejuna and ilea were soaked through a graded series of ethanol and xylene, embedded in paraffin, and sectioned at 5 μm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, USA). The sections were deparaffinized with xylene and rehydrated through graded dilutions of ethanol, and stained with hematoxylin and eosin. The images of jejuna and ilea were acquired using an Olympus simon-01 microscope (Olympus Optical Co., Ltd., Beijing, China). The values of villus height (VH) and crypt depth (CD) were measured 5 times from different villus and crypts per section from each broiler using the Image-Pro Plus software 6.0 (Media Cybernetics, Inc., Washington, USA). VCR was defined as villus height-to-crypt depth ratio.
5
Histological Analysis of Thymus Tissue
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After fixation in 4% PFA for 24 h, the thymus was soaked through a graded series of ethanol and xylene, embedded in paraffin, and sectioned at 5 μm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL). The sections were deparaffinized with xylene and rehydrated through a graded ethanol, and stained with hematoxylin and eosin (HE ). The images were acquired using an Olympus simon-01 microscope (Olympus Optical Co., Ltd., Beijing, China).
6
Jejunal Histomorphometry in Broilers
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With the broiler anesthetized, the abdominal cavity was opened and the jejunum was taken out and the midpoint was found. Approximately 1 cm of the middle jejunum was collected from the chickens 24 h after the last LPS injection at 29 days of age. After fixation in 4% paraformaldehyde for 24 h, the jejunal sections were soaked through a graded series of ethanol and xylene and embedded in paraffin. The jejunum was then sectioned at 5 mm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL, USA). The sections were deparaffinized with xylene and rehydrated through a graded dilution of ethanol. Hematoxylin and eosin (H&E) staining. The jejunal villus height (VH) and crypt depth (CD) were measured according to the method described by Li [13 (link)], and the ratio of VH to CD was also determined. Briefly, eight intact intestinal villi with a straight direction were selected from each sample, and then the VH and CD were investigated. Villus height was defined as the distance from the villus tip to the crypt mouth, and crypt depth was the depth of the distance from the crypt mouth to the base.
7
Intestinal Histomorphometry Analysis
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Fixed intestinal samples were dehydrated using graded ethanol, and vitrificated by dimethylbenzene. Then samples were embedded in paraffin, sliced with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL, USA), and stained by hematoxylin-eosin (HE). The villus height (VH) and crypt depth (CD) were measured using a microscope imaging system (DM 1000, Leica, Germany) and Image-Pro Plus software 6.0 (Media Cybernetics, Inc., Washington, DC, USA) to calculate the ratio of the villus height to the crypt depth (VH/CD).
8
Histological Analysis of Chicken Jejunum
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After fixation in 4% paraformaldehyde for 24 h, the jejunal sections were soaked through a graded series of ethanol and xylene and embedded in paraffin. The jejunum was then sectioned at 5 μm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL). The sections were deparaffinized with xylene and rehydrated through a graded dilution of ethanol. Hematoxylin and eosin (H&E) staining and Alcian Blue-periodic acid Schiff (AB-PAS) staining were performed, respectively. The images were acquired using an Olympus microscope (Olympus Optical Co., Ltd., Beijing, China). The values of villus height (VH), crypt depth (CD), and the number of jejunal goblet cells were measured 6 times from different villus and crypts per section from each chicken using Image-Pro Plus software 6.0 (Media Cybernetics, Rockville, MD, USA). The data were analyzed using 8 sampled chickens per group (n = 8).
9
Placental Histology Analysis
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After fixation in 4% paraformaldehyde for 24 h, the placentas were dehydrated through a graded series of ethanol and xylene, embedded in paraffin wax, and serially sectioned at 5 μm with a Lecia RM2235 microtome (Leica Biosystems Inc., Buffalo Grove, IL, USA). The sections were stained with hematoxylin and eosin (H&E), and viewed under an Olympus simon-01 microscope (Olympus Optical Co., Ltd., Beijing, China). The area of blood sinusoids in the placenta was analyzed using Image-Pro Plus software 6.0 (Media Cybernetics, Inc., Washington, DC, USA).
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