TY  - JOUR
A1  - Endesfelder, Ulrike
A1  - Malkusch, Sebastian
A1  - Flottmann, Benjamin
A1  - Mondry, Justine
A1  - Liguzinski, Piotr
A1  - Verveer, Peter J.
A1  - Heilemann, Mike
T1  - Chemically Induced Photoswitching of Fluorescent Probes - A General Concept for Super-Resolution Microscopy
N2  - We review fluorescent probes that can be photoswitched or photoactivated and are suited for single-molecule localization based super-resolution microscopy. We exploit the underlying photochemical mechanisms that allow photoswitching of many synthetic organic fluorophores in the presence of reducing agents, and study the impact of these on the photoswitching properties of various photoactivatable or photoconvertible fluorescent proteins. We have identified mEos2 as a fluorescent protein that exhibits reversible photoswitching under various imaging buffer conditions and present strategies to characterize reversible photoswitching. Finally, we discuss opportunities to combine fluorescent proteins with organic fluorophores for dual-color photoswitching microscopy.
KW  - Super-Resolution Microscopy
KW  - photoswitchable organic fluorophores
KW  - fluorescent proteins
KW  - super-resolution
KW  - PALM
KW  - dSTORM
Y1  - 2011
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-74896
ER  - 
TY  - JOUR
A1  - Endesfelder, Ulrike
A1  - Malkusch, Sebastian
A1  - Flottmann, Benjamin
A1  - Mondry, Justine
A1  - Liguzinski, Piotr
A1  - Verveer, Peter J.
A1  - Heilemann, Mike
T1  - Chemically Induced Photoswitching of Fluorescent Probes - A General Concept for Super-Resolution Microscopy
JF  - Molecules
N2  - We review fluorescent probes that can be photoswitched or photoactivated and are suited for single-molecule localization based super-resolution microscopy. We exploit the underlying photochemical mechanisms that allow photoswitching of many synthetic organic fluorophores in the presence of reducing agents, and study the impact of these on the photoswitching properties of various photoactivatable or photoconvertible fluorescent proteins. We have identified mEos2 as a fluorescent protein that exhibits reversible photoswitching under various imaging buffer conditions and present strategies to characterize reversible photoswitching. Finally, we discuss opportunities to combine fluorescent proteins with organic fluorophores for dual-color photoswitching microscopy.
KW  - Photoactivated localization microscopy
KW  - Fusion proteins
KW  - Molecules
KW  - Patterns
KW  - Switch
KW  - Limit
KW  - Time
KW  - photoswitchable organic fluorophores
KW  - fluorescent proteins
KW  - super-resolution
KW  - PALM
KW  - dSTORM
Y1  - 2011
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-134080
VL  - 16
IS  - 4
ER  - 
TY  - JOUR
A1  - Wolter, Steve
A1  - Endesfelder, Ulrike
A1  - Linde, Sebastian van de
A1  - Heilemann, Mike
A1  - Sauer, Markus
T1  - Measuring localization performance of super-resolution algorithms on very active samples
JF  - Optics Express
N2  - Super-resolution fluorescence imaging based on  inglemolecule localization relies critically on the availability of efficient processing algorithms to distinguish, identify, and localize emissions of single fluorophores. In multiple current applications, such as threedimensional, time-resolved or cluster imaging, high densities of fluorophore emissions are common. Here, we provide an analytic tool to test the performance and quality of localization microscopy algorithms and demonstrate that common algorithms encounter difficulties for samples with high fluorophore density. We demonstrate that, for typical single-molecule localization microscopy methods such as dSTORM and the commonly used rapidSTORM scheme, computational precision limits the acceptable density of concurrently active fluorophores to 0.6 per square micrometer and that the number of successfully localized fluorophores per frame is limited to 0.2 per square micrometer.
Y1  - 2011
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-85936
ER  -