University of Hertfordshire

From the same journal

By the same authors

  • Dan Stratton
  • Sigrun Lange
  • Sharad Kholia
  • Samireh Jorfi
  • Samuel Antwi-Baffour
  • Jameel Inal
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Original languageEnglish
Number of pages6
Pages (from-to)619-624
JournalBiochemical and Biophysical Research Communications
Journal publication date24 Oct 2014
Volume453
Issue3
Early online date6 Oct 2014
DOIs
Publication statusPublished - 24 Oct 2014

Abstract

Using a Quartz Crystal Microbalance with dissipation monitoring, QCM-D (label-free system) measuring changes in resonant frequency (Δf) that equate to mass deposited on a sensor, we showed the attachment, over a 60 min period, of a monolayer of PC3 cells to the gold electrodes of the quartz crystal sensor, which had been rendered hydrophilic. That MVs were released upon BzATP stimulation of cells was confirmed by NTA analysis (average 250 nm diameter), flow cytometry, showing high phosphatidylserine exposition and by fluorescent (Annexin V Alexa Fluor® 488-positive) and electron microscopy. Over a period of 1000s (16.7 min) during which early apoptosis increased from 4% plateauing at 10% and late apoptosis rose to 2%, the Δf increased 20 Hz, thereupon remaining constant for the last 1000s of the experiment. Using the Sauerbrey equation, the loss in mass, which corresponded to the release of 2.36 × 106 MVs, was calculated to be 23 ng. We therefore estimated the mass of an MV to be 0.24 pg. With the deposition on the QCM-D of 3.5 × 107 MVs over 200s, the decrease in Δf (Hz) gave an estimate of 0.235 pg per MV.

ID: 13068971