Mice were deeply anesthetized and perfused intracardially with 4% paraformaldehyde (PFA) in 0.1 M phosphate buffered saline. The femora were removed and then post-fixed for 48 h in the same fixative at 4 °C. After demineralization in EDTA (10%) for 10 days, femur samples were dehydrated in an ascending gradient of ethanol (30–100%) followed by paraffin embedding. Serial sections for trabecular bone were obtained from the distal femur at a thickness of 5 µm followed by tartrate-resistant acid phosphatase (TRAP) staining or alkaline phosphatase (ALP) staining using TRAP kits (Fast Red TR/Naphthol AS-MX, Sigma, St. Louis, MO) and NBT/BCIP (Thermo Scientific), respectively30 (link). Bone static histomorphometric analyses for osteoclast number (osteoclast number per trabecular bone surface covered by osteoclasts, Oc.S/BS) and osteoblast number (osteoblast number per trabecular bone surface, Ob.N/BS) were conducted using Image J (NIH) based on images taken by a Leica Q500MC microscope. Osteoclasts, osteoblasts and trabecular bone at the metaphysis of the femur (1500 µm proximal to the distal growth plate) were quantified, since bone destruction in this model mainly occurs in this area36 (link),65 (link). Three sections per animal were randomly chosen and used for quantification.
Q500mc microscope
Manufactured by Leica
Sourced in Spain, Germany
The Leica Q500MC is a digital microscope designed for laboratory use. It features a high-resolution camera and advanced optics to capture detailed images of samples. The microscope can be connected to a computer for image analysis and documentation.
Automatically generated - may contain errors
6 protocols using q500mc microscope
1
Histomorphometric Analysis of Bone
2
Lung Histology: Tissue Fixation and Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were deeply anesthetized with isoflurane and perfused intracardially with PBS, followed by 4% PFA. After the perfusion, lungs were removed from mice and post-fixed in the same fixative overnight. The samples were then dehydrated with a 30% sucrose solution, embedded in O.C.T. (Tissue Tek), and cryosectioned to produce 8 μm thick sections. For Hematoxylin and Eosin (H & E) staining, lung sections were rehydrated and stained with 0.1% Hematoxylin and 0.5% Eosin in sequence. After dehydration and clearance with HistoClear (Electron Microscopy Sciences, Hatfield, PA), slides were mounted with a resinous mounting medium (Mercedes Medical, Sarasota, FL) and subsequently imaged using a Leica Q500MC microscope with a digital camera at different magnifications.
3
Histological Analysis of Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were deeply anesthetized with isoflurane and perfused intracardially with PBS, followed by 4% PFA. After the perfusion, lungs were removed from mice and post-fixed in the same fixative overnight. The samples were then dehydrated with a 30% sucrose solution, embedded in O.C.T.
(Tissue Tek), and cryosectioned to produce 8 μm thick sections. For Hematoxylin and Eosin (H & E) staining, lung sections were rehydrated and stained with 0.1% Hematoxylin and 0.5% Eosin in sequence. After dehydration and clearance with HistoClear (Electron Microscopy Sciences, Hatfield, PA), slides were mounted with a resinous mounting medium (Mercedes Medical, Sarasota, FL) and subsequently imaged using a Leica Q500MC microscope with a digital camera at different magnifications.
(Tissue Tek), and cryosectioned to produce 8 μm thick sections. For Hematoxylin and Eosin (H & E) staining, lung sections were rehydrated and stained with 0.1% Hematoxylin and 0.5% Eosin in sequence. After dehydration and clearance with HistoClear (Electron Microscopy Sciences, Hatfield, PA), slides were mounted with a resinous mounting medium (Mercedes Medical, Sarasota, FL) and subsequently imaged using a Leica Q500MC microscope with a digital camera at different magnifications.
4
Quantifying Osteoclasts and Osteoblasts in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were deeply anesthetized and perfused intracardially with 4% paraformaldehyde (PFA) in 0.1M phosphate buffered saline. The femora were removed and then post-fixed for 48h in the same fixative at 4 °C. After demineralization in EDTA (10%) for 10 days, femur samples were dehydrated in an ascending gradient of ethanol (30-100%) followed by paraffin embedding. Serial sections for trabecular bone were obtained from the distal femur at a thickness of 5 µm followed by tartrate-resistant acid phosphatase (TRAP) staining or alkaline phosphatase (ALP) staining using TRAP kits (Fast Red TR/Naphthol AS-MX, Sigma, St. Louis, MO) and NBT/BCIP (Thermo Scientific), respectively 27 (link) . Bone static histomorphometric analyses for osteoclast number (osteoclast number per trabecular bone surface covered by osteoclasts, Oc.S/BS) and osteoblast number (osteoblast number per trabecular bone surface, Ob.N/BS) were conducted using Image J (NIH) based on images taken by a Leica Q500MC microscope. Osteoclasts, osteoblasts and trabecular bone at the metaphysis of the femur (1500 µm proximal to the distal growth plate) were quantified, since bone destruction in this model mainly occurs in this area 33, (link)51 (link) . Three sections per animal were randomly chosen and used for quantification.
5
Quantitative Histomorphometry of Implants
Check if the same lab product or an alternative is used in the 5 most similar protocols
The digital quantitative analysis was performed by using calibrated digital images, ranging from 4
×to 40
×magnification (Leica microscope Q500Mc, Leica DFC320s, 3,088 × 2,550 pixels, Leica Microsystems, Barcelona, Spain). The most central sagittal section of each implant was taken for histomorphometric analysis using MIP 4.5 software (Microms Image Processing Software, CID, Consulting Image Digital, Barcelona, Spain), and connected to a Sony DXC-151s 2/3-CCD RGB Color Video Camera. The areas of interest were marked, and their values were calculated digitally for the total percentage with the ImageJ software (W. Rasband, National Institutes of Health, Maryland, United States). The evaluation consisted of the measurements of new bone formation (NB), residual biomaterial (RB), and CT in relation to the total measurement area. Values were expressed in percentage.
+ Expand
The digital quantitative analysis was performed by using calibrated digital images, ranging from 4
×to 40
×magnification (Leica microscope Q500Mc, Leica DFC320s, 3,088 × 2,550 pixels, Leica Microsystems, Barcelona, Spain). The most central sagittal section of each implant was taken for histomorphometric analysis using MIP 4.5 software (Microms Image Processing Software, CID, Consulting Image Digital, Barcelona, Spain), and connected to a Sony DXC-151s 2/3-CCD RGB Color Video Camera. The areas of interest were marked, and their values were calculated digitally for the total percentage with the ImageJ software (W. Rasband, National Institutes of Health, Maryland, United States). The evaluation consisted of the measurements of new bone formation (NB), residual biomaterial (RB), and CT in relation to the total measurement area. Values were expressed in percentage.
6
Histomorphometric Analysis of Implants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histomorphometric analysis was performed using calibrated digital images at ×10 magnification (Leica microscope Q500Mc, Leica DFC320s, 3088 × 2550 pixels, Leica Microsystems, Barcelona, Germany). The most central sagittal section of each implant was taken for the histomorphometric analysis using MIP 4.5 software (Microns Image Processing Software, CID, Consulting Image Digital, Barcelona, Spain) connected to a Sony DXC-151s 2/3-CCD RGB Color Video Camera.
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!