We assessed osteoblast and osteoclast activity in response to microgravity and BP-NELL-PEG treatment via semi-quantitation of staining for TRAP, osteocalcin, and RANKL in mouse femurs. Formalin-fixed paraffin-embedded sections of mouse bones (5 μm thick) from femur were cut with a microtome and mounted on charged slides (n = 3 per group). Sections were either subjected to standard hematoxylin-eosin (H&E) staining for overview or TRAP (Sigma-Aldrich, Cat# 387A-1KT) staining, and immunolabeled with the following antibodies: osteocalcin (Santa Cruz, Cat# sc-365797) and RANKL (Abcam, Cat# ab45039). Photomicrographs were acquired using Olympus BX 51 and IX 71 microscopes equipped with the Cell Sense digital imaging system (Olympus, Japan). To assess immunohistochemistry, digitized photographs were acquired and the intensity of positive staining was automatically measured using ImageJ 1.45 (National Institute of Health, freeware imaging software, https://imagej.nih.gov/nih-image/ ).
Cell sense digital imaging system
Manufactured by Olympus
Sourced in Japan
The Cell Sense digital imaging system is a compact, high-performance microscope imaging system designed for life science applications. It features advanced image acquisition and processing capabilities to capture detailed images of cells and other small biological samples.
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Lab products found in correlation
2 protocols using cell sense digital imaging system
1
Assessing Osteoblast and Osteoclast Activity
2
Characterizing Muscle Fiber Composition
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Formalin-fixed paraffin-embedded sections of mouse muscle (5um thickness) from gastrocnemius and quadriceps were cut with a microtome and mounted on charged slides. Sections were either subjected to standard hematoxylin-eosin (H&E) staining for overview, or immunolabeled with the following antimyosin heavy chain (MyHC) isoform antibodies: type I MyHC isoform (Abcam, Cat# ab11083); type II MyHC isoform (Abcam, Cat# ab51263). The sections were co-stained with an anti-laminin antibody (Abcam, Cat# ab11575) to allow measurement of fiber size. In all protocols, donkey anti rabbit Alexa-488 conjugated secondary antibody was used for laminin staining, donkey anti-mouse Alexa-594 conjugated secondary antibody for type I MyHC antigen staining, and anti-mouse Alexa-488 conjugated secondary antibody for type II MyHC antigen staining. Photomicrographs were acquired using Olympus BX 51 and IX 71 microscopes equipped with Cell Sense digital imaging system (Olympus, Japan). To assess the cross-sectional area (CSA) of the different myofiber types, digitized photographs were acquired and the myofiber CSA was automatically measured by means of ImageJ 1.45 g (NIH, freeware imaging software).
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