正帯電金ナノ粒子標識法による鞭毛・線毛・細胞突起の大気圧走査電子顕微鏡観察  [in Japanese] Atmospheric Scanning Electron Microscope Observation of Cells Using a Charged Gold Labeling Technique  [in Japanese]

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Abstract

<p>細菌や培養細胞の光学顕微鏡観察には分解能の限界があり,ナノメートルオーダーの細菌の鞭毛・線毛や,真核生物細胞のフィロポディアの形状観察は容易ではない.このような繊細な構造を電子顕微鏡で観察するには,脱水や蒸着等の繁雑な前処理が必要であり,これによって本来の形状が失われる場合もある.本研究では細胞表面が負帯電していることを利用して,正帯電金ナノ粒子を細胞に吸着させた後,金増感を行い水溶液中で大気圧走査電子顕微鏡(ASEM)観察を試みたところ,サルモネラ菌のらせん状の鞭毛,大腸菌の線毛,COS7細胞のフィロポディアを明確に観察することができた.観察に必要な前処理は水溶液を入れ替えるだけの簡単な操作であり,1時間ほどで完了可能であった.本稿で紹介した技術は細胞本来の微細な形状を容易に観察することができるため,生物分野に広く応用が期待される.</p>

<p>Optical and electron microscopes are widely used for morphological observation of microbes and cultured cells. However, the small size of microbes (flagella and pili) and cells (filopodia) limits optical microscope observation. An electron microscope has a high resolution. However, lengthy pretreatments such as dehydration or metal evaporation are basically necessary. This may obscure delicate structures in SEM images. For the observation of natural structures in microbes and cells, we have developed an in-liquid gold labeling technique using an atmospheric SEM (ASEM), which observes a sample in liquid in open atmosphere. Sample pretreatment required only simple tasks including fixation, gold labeling, and medium exchange, taking less than one hour in total. ASEM imaging of the bacteria in aqueous solution revealed pili and delicate spiral flagella in natural form. The gold labeling also allowed filopodia as thin as 100 nm in diameter to be clearly visualized.</p>

Journal

  • KENBIKYO

    KENBIKYO 48(3), 190-194, 2013

    The Japanese Society of Microscopy

Codes

  • NII Article ID (NAID)
    130007705268
  • NII NACSIS-CAT ID (NCID)
    AA11917781
  • Text Lang
    JPN
  • ISSN
    1349-0958
  • NDL Article ID
    025156148
  • NDL Call No.
    Z16-896
  • Data Source
    NDL  J-STAGE 
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