Detection of mRNA in single living cells using AFM nanoprobes.
Examining messenger RNA (mRNA) expression is useful for the determination of cell and tissue conditions. Many methods of determining mRNA expression require total RNA extraction or cell fixation. These processes cause difficulties in examining mRNA expression in single living cells without causing cell death. We think that analysis of specific mRNA expression in single living cells will become important in cell biology. In this chapter, we present a method to examine mRNA expression of single living cells without killing the cells. The single-cell nanoprobe (SCN) method uses atomic force microscopy (AFM) to extract mRNA. We also present examples of beta-actin mRNA detection and multiple mRNA detection from single living cells.
Uehara
Hironori
H
Department of Ophthalmology and Visual Science, University of Utah, Salt Lake City, UT 84132, USA. Hironori.Uehara@hsc.utah.edu
Ikai
Atsushi
A
Osada
Toshiya
T
D000199:Actins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002477:Cells / Q000378:metabolism*; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D018625:Microscopy, Atomic Force / Q000379:methods*; D049329:Nanostructures; D036103:Nanotechnology; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019147:Vomeronasal Organ / Q000166:cytology / Q000378:metabolism
D000199:Actins; D017931:DNA Primers; D012333:RNA, Messenger