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KMID : 1101220030350020196
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2003 Volume.35 No. 2 p.196 ~ p.199
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Comparative Analysis of Skeletal Muscle Snap Freezing Method For Examination of Cellular Morphology in Differentiation of Myogenic or Neurogenic Disease
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Han Jang-Hyun
Chung Hak-In
Cho Myung-Hwan
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Abstract
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Muscle biopsy is a critical step in caring patients having variable myopathies. The differential diagnosis of neurogenic or myogenic origin is very important. Variable artifacts such as ice crystals can interfere with the exact diagnosis. To acquire an optimal section, we compared several frozen and sectioning methods. To avoid direct contact, a piece of cork covered with water or an embedded medium is used as a moving plate for muscle biopsy specimens. During the indirect method, the cork should be dropped into the gel or between liquid and solid state of isopentane cooled by liquid nitrogen using a forceps. The sample must be kept in a -70℃ ultra low freezer untill sectioning. Prior to sectioning, leave the sample in cryostat for 20 minutes minimally in order to recover the optimal temperature. Sectioning must be done in a cryostat cooled at -20℃ with a new disposal knife at 7 to 10 μm for prevention of cracking. At this time, sectioning without OCT compound embedding can avoid the ice crystal artifact. Methanol prefix should be done 3 to 5 seconds after sectioning. The muscle biopsy sample must be handled with minimal trauma and precautions in order to acquire optimal sections for direct diagnosis.
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KEYWORD
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Ice crystals, Cork, Isopentane cooled by liquid nitrogen, Prefix
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