Living cell positioning on the surface of gold film for SPR analysis

Abstract

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 HydroCell/simple pipetting, with RepCell/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, on RepCell, 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 or RepCell, suggesting that trypsin may harm molecules involved in cellular reactions. Thus, the methods of cell fixation and removal with HydroCell or RepCell should enable us to analyze various reactions in either adherent or non-adherent cells by SPR.

Surface plasmon resonance sensors in cell biology: basics and application

“…Based on two of the above-mentioned reports, Yanase et al compared different methods to fix living cells on a SPR gold surface and to recover living and proliferating adherent cells, that could not be directly cultured on the surface, from the culture dish [93]. As sensoric elements, the group used two different types of cells: as non-adherent cell type, human basophils and B-cells were fixed to the sensor chip either by a biocompatible anchor for cell membranes, an aminoalkanethiol or an amino-reactive cross-linker.…”

“…The different series of experiments conducted in this study identified aminoalkanethiols, like cysteamine, as the linkers of choice when it comes to an easily applicable and equally efficient surface immobilization of non-adherent cells. The transfer and recovery of adherent cells that are not directly grown on the sensor surface but on a culture dish and subsequently transferred to the sensor, works best with special culture dish materials fabricated from superhydrophile property or temperature-responsive polymers, that keep adherent cells in a quasi-floating state and make them easily removable by simple pipetting [93].…”

The effect of electrical stimulation on the differentiation of hESCs adhered onto fibronectin-coated gold nanoparticles

“…Gold has been the most frequently investigated conductive material due to its ability to easily convey electrical shock to substrates during electrical charge stimulation of affected cells or tissues [5][6][7]. In biomedical fields, conductive gold films have been reported to enhance the proliferation and differentiation of adhered cells such as nerve and endothelial cells [8][9][10]. Thus, gold-coated surfaces have suitable biocompatibility, which indicates that they have the potential for biomedical applications [11][12][13].…”

Kinetic studies of bovine serum albumin interaction with PG and TBHQ using surface plasmon resonance

“…In addition to these procedures, surface plasmon resonance (SPR) technique has been developed during recent two decades for investigation of interaction between two ingredients like antigen-antibody [18], cell-drug [19], drug-protein [20][21][22], DNA-RNA and etc [23]. Also calculation of reliable affinity and kinetics binding constants like association rate constant (ka), dissociation rate constant (kd) and affinity (KD) has been reported using SPR [24][25][26].…”