Nanostructure analysis using spatially modulated illumination microscopy.
We describe the usage of the spatially modulated illumination (SMI) microscope to estimate the sizes (and/or positions) of fluorescently labeled cellular nanostructures, including a brief introduction to the instrument and its handling. The principle setup of the SMI microscope will be introduced to explain the measures necessary for a successful nanostructure analysis, before the steps for sample preparation, data acquisition and evaluation are given. The protocol starts with cells already attached to the cover glass. The protocol and duration outlined here are typical for fixed specimens; however, considerably faster data acquisition and in vivo measurements are possible.
Baddeley
David
D
Kirchhoff Institut für Physik, Universität Heidelberg, INF 227, D-69120 Heidelberg, Germany. baddeley@kip.uni-heidelberg.de
Batram
Claudia
C
Weiland
Yanina
Y
Cremer
Christoph
C
Birk
Udo J
UJ
D002138:Calibration; D002477:Cells / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D049329:Nanostructures / Q000032:analysis*