Living cell positioning on the surface of gold film for SPR analysis
スポンサーリンク
概要
- 論文の詳細を見る
Living cell reactions are detected as changes of the angle of resonance (AR) for surface plasmon resonance (SPR). Since SPR reflects the events in the field of evanescence, cells need to be fixed on the sensor chip. In this study, we developed methods to fix living cells on a gold surface and to recover adherent cells from the culture dish, preserving their functions to be analyzed by SPR. Human basophils and B cells were fixed to the sensor chip by a biocompatible anchor for cell membranes (alpha-succinimidyloxysuccinyl omega-oleyloxy polyoxyethylene), aminoalkanethiol (cyteamine, 8-amino octanethiol) or an amino-reactive cross-linker (dithiobis [succinimidylpropionate]). They showed an increase of AR in response to various stimuli. RBL-2H3 cells, which firmly adhered to the culture dish, were cultured/recovered with HydroCelI(TM)/Simple pipetting, with RepCelI(TM)/pipetting at 4 degrees C, or on normal plastic culture dishes with trypsinization or by scraping at 4 degrees C, respectively. The exocytosis of RBL-2H3 cells was largely impaired by scraping, but only slightly by the treatment with pipetting on HydroCell(TM), on RepCell(TM) or with trypsin. The membrane ruffling of the cells prepared by the last three treatments induced by antigens appeared the same. However, the change of AR with cells prepared by trypsin and those by scraping at 4 degrees C were lower than those by HydroCell(TM) or RepCell(TM), suggesting that trypsin may harm molecules involved in cellular reactions. Thus, the methods of cell fixation and removal with HydroCell(TM) or RepCell(TM) should enable us to analyze various reactions in either adherent or non-adherent cells by SPR.
論文 | ランダム
- 本当に「闘う政治家」なのか (安倍新内閣を採点する)
- 将来計画と事業仕分け
- 規制強化に対応するためキャッシング・ビジネスをどう改革するか--コスト削減と残高減少の緩和を柱に、総合的な施策を打ち出すべし (特集 徹底検証!! 貸金業制度改革の影響と課題)
- 「闘う政治家」を支える行動派シンクタンク
- 新入園児の適応過程について--情緒的対象恒常性の視点から