Resorption pit areas were measured from Field Emission Scanning Electron Microscopy (FESEM) images with Merz grid analysis. After multinuclear cell counting, cells were detached from the slices by brushing, samples were dehydrated in ascending ethanol series and dried with a critical point drying equipment K850 (Quorum technologies, UK). Samples were coated with 5 nm platinum by Q150T ES sputter coater (Quorum Technologies) and viewed with Sigma HD VP FE-SEM (Carl Zeiss Microscopy GmbH, Germany). FESEM images were taken from three fields (voltage 5.0 kV, magnification 50×, area 0,035 cm2) from each bone slice, n = 3. The morphometric analysis of the pits was performed with ImageJ 1.49t software (NIH, USA) by superimposing a Merz grid with 80–88 points in semicircular lines over the image. Points in pits were counted and the proportion of resorption pits versus intact bone surface was counted. The proportion of resorbed area was normalized to multinuclear cell number (counting described above), and the average area resorbed by one cell was calculated.
Sigma hd vp fe sem
Manufactured by Zeiss
Sourced in Germany, United Kingdom
The Sigma HD VP FE-SEM is a field emission scanning electron microscope (FE-SEM) designed for high-resolution imaging and analysis. It features a high-brightness electron source, advanced optics, and a versatile vacuum system to enable detailed examination of a wide range of sample types.
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Lab products found in correlation
5 protocols using sigma hd vp fe sem
1
Quantifying Osteoclast Resorption Activity
2
Quantifying Bone Resorption via FESEM Imaging
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Resorption pit areas were measured from Field Emission Scanning Electron Microscopy (FESEM) images with Merz grid analysis. After multinuclear cell counting, cells were detached from the slices by brushing, samples were dehydrated in ascending ethanol series and dried with a critical point drying equipment K850 (Quorum technologies, UK). Samples were coated with 5 nm platinum by Q150T ES sputter coater (Quorum Technologies) and viewed with Sigma HD VP FE-SEM (Carl Zeiss Microscopy GmbH, Germany). FESEM images were taken from three fields (voltage 5.0 kV, magnification 50 ×, area 0.039 cm2) from each bone slice, n = 3. The morphometric analysis of the pits was performed with ImageJ 1.51n software (NIH, USA) by superimposing a Merz grid with 92–96 points in semicircular lines over the image. Points in pits were counted and the percentage of resorption pits versus intact bone surface was counted. The percentage of resorbed area was normalized to multinuclear cell number (counting described above), and the average area resorbed by one cell was calculated.
3
Critical Point Drying and SEM Imaging
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The ALD-HA samples were dehydrated in ascending ethanol series and dried with a critical point drying equipment K850 (Quorum Technologies, Lewes, UK). Samples were coated with 5 nm platinum by Q150T ES sputter coater (Quorum Technologies, Lewes, UK) and viewed with Sigma HD VP FE-SEM (Carl Zeiss Microscopy GmbH, Oberkochen, Germany). FESEM images were taken with 5.0 kV voltage.
4
Quantifying Bone Resorption via FESEM
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Resorption pit areas were compared from Field Emission Scanning Electron Microscopy (FESEM) images with Merz grid analysis. Cells were wiped away from the bone slices. Samples were dehydrated in an ascending ethanol series and dried with critical point drying equipment K850 (Quorum technologies, UK). The samples were sputter coated with 5 nm platinum with a Q150T ES sputter coater (Quorum Technologies) and viewed with a Sigma HD VP FE-SEM (Carl Zeiss Microscopy GmbH, Germany). FESEM images were taken from three fields (voltage 5.0 kV, magnification 50x, area 0.035 cm2) from each bone slice, n = 3. The morphometric analysis of the resorption pits was performed with ImageJ 1.49t software (NIH, USA) by superimposing a Merz grid over the image. 80–88 points from semicircular lines were counted and the proportion of resorption pits versus intact bone surface was determined.
5
Critical Point Drying and SEM Imaging
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The bone slices, non-coated Ti samples, and ALD-HA samples were dehydrated in ascending ethanol series and dried with a critical point drying equipment K850 (Quorum Technologies, Lewes, UK). Samples were coated with 5 nm platinum by Q150T ES sputter coater (Quorum Technologies, Lewes, UK) and viewed with Sigma HD VP FE-SEM (Carl Zeiss Microscopy GmbH, Oberkochen, Germany). FESEM images were taken with 5.0 kV voltage.
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