このエントリーをはてなブックマークに追加
ID 21470
本文ファイル
著者
Yanase, Yuhki
Suzuki, Hidenori
Tsutsui, Tomoko
Uechi, Ichiro
Mihara, Shoji
キーワード
biosensor
surface plasmon resonance
non-adherent cells
cell fixation
recovery
NDC
医学
抄録(英)
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.
掲載誌名
Biosensors & Bioelectronics
23巻
4号
開始ページ
562
終了ページ
567
出版年月日
2007-11
出版者
Elsevier Advanced Technology
ISSN
0956-5663
NCID
出版者DOI
言語
英語
NII資源タイプ
学術雑誌論文
広大資料タイプ
学術雑誌論文
DCMIタイプ
text
フォーマット
application/pdf
著者版フラグ
author
権利情報
Copyright(c) 2007 Elsevier Advanced Technology
関連情報URL(IsVersionOf)
http://dx.doi.org/10.1016/j.bios.2007.07.005
部局名
医歯薬学総合研究科