• Title/Summary/Keyword: HVEM

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Whole Mount Preparation of Primary Cultured Neuron for HVEM Observation (배양된 시경세포 관찰을 위한 초고압전자현미경 홀마운트 시료제작기법)

  • Kim, Hyun-Wook;Hong, Soon-Taek;Oh, Seung-Hak;Park, Chang-Hyun;Kim, Hyun;Rhyu, Im-Joo
    • Applied Microscopy
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    • v.41 no.1
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    • pp.69-73
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    • 2011

  • High-voltage electron microscope (HVEM) has higher resolution and penetration power than conventional transmission electron microscope that could be load thick specimen. Some researchers have taken this advantage of HVEM to explore 3-dimensional configuration of the biological structures including tissue and cells. Whole mount preparations has been employed to study some cell lines and primary culture cells. In this study, we would like to introduce useful whole mount preparation method for neuronal studies. The plastic coverslips were punched, covered by formvar membrane and coated with carbon. The neurons obtained embryonic 18 rat hippocampus were seeded on the prepared cover slip. The coverslips were fixed, dried in freeze drier and kept in a descicator until HVEM observation. We could observe detailed neuronal structures such as soma, dendrite and spine under HVEM without conventional thin section and heavy metal stain. The anaglyphic image based on stereo paired image ($-8^{\circ},+8^{\circ}$) provides three dimensional perception of the neuronal dendrites and their spines. This method could be applied to sophisticated analysis of dendritic spine under the various experimental conditions.

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High Voltage Electron Microscopy of Structural Patterns of Plastid Crystalline Bodies in Sedum rotundifolium (HVEM에 의한 둥근잎꿩의 비름 (Sedum rotundifolium L.) 색소체의 결정체 구조)

  • Kim, In-Sun
    • Applied Microscopy
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    • v.36 no.2
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    • pp.73-82
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    • 2006

  • Major contributions has been made in cellular ultrastructure studies with the use of high voltage electron microscopy (HVEM) and tomography. Applications of HVEM, accompanied by appropriate image processing, have provided great improvements in the analysis of three-dimensional cellular structures. In the present study, structural patterns of the crystalline bodies that are distinguished in mesophyll plastids of CAM-performing Sedum rotundifolium L., have been investigated using HVEM and tomography. Tilting, and diffraction pattern analysis were performed during the investigation. The titlting was performed at ${\pm}60^{\circ}\;with\;2^{\circ}$ increments while examining serial sections ranging from 0.125 to $1{\mu}m$ in thickness. The young plastids exhibited crystalline inclusion bodies that revealed a peculiar structural pattern. They were irregular in shape and also variable in size. Their structural attributes affected the plastid morphology. The body consisted of a large number of tubular elements, often reaching up to several thousand in number. The tubular elements typically aggregated to form a fluster The elements demonstrated either a parallel or lattice arrangement depending on the sectioning angle. The distance between the elements was approximately 20nm as demonstrated by the diffraction analysis. HVEM examination of the serial sections revealed an occasional fusion or branching of elements within the inclusion bodies. Finally, a three-dimensional reconstruction of the plastid crystalline bodies has been attempted using two different image processing methods.

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