Combining confocal and BSE SEM imaging for bone block surfaces.
نویسندگان
چکیده
The present report presents a method for the correlation of qualitative and quantitative BSE SEM imaging with confocal scanning light microscopy (CSLM) imaging modes applied to bone samples embedded in PMMA. The SEM has a proper digital scan generator: we leave the BSE image unchanged, and match the CSLM image to it, because the CSLM scan mechanism is not digital, though the signal is digitised. Our overlapping program uses a linear transformation matrix which projects one system to the other, calculated by finding three corresponding points in BSE and CSLM pictures. BSE images are empty where cells and osteoid are present. Fluorescence mode CSLM fills in these gaps. The combination images enhance our understanding of what is going on - and re-establish the need for good cellular preservation.
منابع مشابه
Staining plastic blocks with triiodide to image cells and soft tissues in backscattered electron SEM of skeletal and dental tissues.
Backscattered electron scanning electron microscopy (BSE SEM) is an invaluable method for studying the histology of the hard, mineralised components of poly-methyl methacrylate (PMMA) or other resin embedded skeletal and dental tissues. Intact tissues are studied in micro-milled or polished block faces with an electron-optical section thickness of the order of a half to one micron and with the ...
متن کاملSelective Oligonucleotide and MRNA Pull-Down with Shielded Covalent Probes
2556-Pos Board B575 Bringing Deconvolution Algorithmic Techniques to the Electron Microscope Ben Lich, Xiaodong Zhuge, Pavel Potocek, Faysal Boughorbel, Cliff Mathisen. FEI Company, Eindhoven, Netherlands. The Scanning Electron Microscope (SEM) as a three dimensional imaging tool is increasingly popular for studies in neurobiology. Serial slicing methods based on diamond-knife cutting are, howe...
متن کاملA New Microstructural Imaging Approach through EBSD Pattern Region of Interest Analysis
The requirements of a highly tilted sample and a well-polished surface for optimal EBSD pattern collection can make traditional SEM imaging challenging. With a tilted sample, SEM SE and BSE detectors are often in non-optimal positions for electron detection. For many materials, the polished surfaces produce weak contrast with SE imaging. Traditionally placed BSE detectors must be used with care...
متن کاملRegistration for Correlative Microscopy Using Image Analogies
Abstract. Correlative microscopy is a methodology combining the functionality of light microscopy with the high resolution of electron microscopy and other microscopy technologies for the same biological specimen. In this paper, we propose an image registration method for correlative microscopy, which is challenging due to the distinct appearance of biological structures when imaged with di↵ere...
متن کاملClarification of the Structure and Bone Mineral Content of Heterotopic Ossification
INTRODUCTION Heterotopic ossification (HO) is a pathological process characterized by ectopic osseous growth in muscle and periarticular regions. Although HO may develop from rare genetic disorders, abnormal bone growth has been most frequently reported following trauma, arthroplasty, burns, spinal cord injury and traumatic brain injury. The amount of HO that forms following soft tissue injury ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- European cells & materials
دوره 9 شماره
صفحات -
تاریخ انتشار 2005