Journal
BIOSENSORS & BIOELECTRONICS
Volume 25, Issue 1, Pages 22-27Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2009.05.036
Keywords
Nanoneedle; AFM; Single cell; FRET; Caspase-3 activity; MOMENT
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Funding
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Research and Development in a New Converting Field Based on Nanotechnology and Materials Sciences to JM [19770163]
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A method to detect an enzymatic reaction in a single living cell using an atomic force microscope equipped with an ultra-thin needle (a nanoneedle) and a fluorescent probe molecule was developed. The nanoneedle enables the low-invasive delivery of molecules attached onto its surface directly into a single cell. We hypothesized that an enzymatic reaction in a cell could be profiled by monitoring a probe immobilized on a nanoneedle introduced into the cell. In this study, a new probe substrate (NHGcas546) for caspase-3 activity based on fluorescent resonance energy transfer (FRET) was constructed and fixed on a nanoneedle. The NHGcas546-modified nanoneedle was inserted into apoptotic cells, in which caspase-3 is activated after apoptosis induction, and a change in the emission spectrum of the immobilized probe could be observed on the surface of the nanoneedle. Thus, we have developed a successful practical method for detecting a biological phenomenon in a single cell. We call the method MOlecular MEter with Nanoneedle Technology (MOMENT). (C) 2009 Elsevier B.V. All rights reserved.
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