TY  - JOUR
A1  - Nanguneri, Siddharth
A1  - Flottmann, Benjamin
A1  - Horstmann, Heinz
A1  - Heilemann, Mike
A1  - Kuner, Thomas
T1  - Three-Dimensional, Tomographic Super-Resolution Fluorescence Imaging of Serially Sectioned Thick Samples
JF  - PLoS One
N2  - Three-dimensional fluorescence imaging of thick tissue samples with near-molecular resolution remains a fundamental challenge in the life sciences. To tackle this, we developed tomoSTORM, an approach combining single-molecule localization-based super-resolution microscopy with array tomography of structurally intact brain tissue. Consecutive sections organized in a ribbon were serially imaged with a lateral resolution of 28 nm and an axial resolution of 40 nm in tissue volumes of up to 50 \(\mu\)mx50\(\mu\)mx2.5\(\mu\)m. Using targeted expression of membrane bound (m)GFP and immunohistochemistry at the calyx of Held, a model synapse for central glutamatergic neurotransmission, we delineated the course of the membrane and fine-structure of mitochondria. This method allows multiplexed super-resolution imaging in large tissue volumes with a resolution three orders of magnitude better than confocal microscopy.
KW  - architecture
KW  - rat calyx
KW  - in-vivo
KW  - microscopy
KW  - resolution
KW  - proteins
KW  - transmission
KW  - ultrastructure
KW  - reconstruction
KW  - localization
Y1  - 2012
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-134434
VL  - 7
IS  - 5
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
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  -