TY  - CHAP
Y1  - 2015///
AV  - restricted
ED  - Paluch, Ewa K
EP  -  117
TI  - High-content 3D multicolor super-resolution
localization microscopy
T3  - Methods in Cell Biology
KW  - 3D; dSTORM; High-content screening; Immunofluorescence; Localization microscopy; Multicolor; Super-resolution
CY  - Amsterdam, The Netherlands
PB  - Elsevier
T2  - Biophysical Methods in Cell Biology
N2  - Super-resolution (SR) methodologies permit the visualization of cellular structures at near-molecular scale (1?30 nm), enabling novel mechanistic analysis of key events in cell biology not resolvable by conventional fluorescence imaging (?300-nm resolution). When this level of detail is combined with computing power and fast and reliable analysis software, high-content screenings using SR becomes a practical option to address multiple biological questions. The importance of combining these powerful analytical techniques cannot be ignored, as they can address phenotypic changes on the molecular scale and in a statistically robust manner. In this work, we suggest an easy-to-implement protocol that can be applied to set up a high-content 3D SR experiment with user-friendly and freely available software. The protocol can be divided into two main parts: chamber and sample preparation, where a protocol to set up a direct STORM (dSTORM) sample is presented; and a second part where a protocol for image acquisition and analysis is described. We intend to take the reader step-by-step through the experimental process highlighting possible experimental bottlenecks and possible improvements based on recent developments in the field.
ID  - discovery1467957
N1  - This version is the version of record. For information on re-use, please refer to the publisher?s terms and conditions.
SP  - 95 
A1  - Pereira, Pedro M
A1  - Almada, Pedro
A1  - Henriques, Ricardo
M1  - 125
UR  - http://dx.doi.org/10.1016/bs.mcb.2014.10.004
SN  - 0091-679X
ER  -