TY  - JOUR
A1  - Markert, Sebastian Matthias
A1  - Britz, Sebastian
A1  - Proppert, Sven
A1  - Lang, Marietta
A1  - Witvliet, Daniel
A1  - Mulcahy, Ben
A1  - Sauer, Markus
A1  - Zhen, Mei
A1  - Bessereau, Jean-Louis
A1  - Stigloher, Christian
T1  - Filling the gap: adding super-resolution to array tomography for correlated ultrastructural and molecular identification of electrical synapses at the C. elegans connectome
JF  - Neurophotonics
N2  - Correlating molecular labeling at the ultrastructural level with high confidence remains challenging. Array tomography (AT) allows for a combination of fluorescence and electron microscopy (EM) to visualize subcellular protein localization on serial EM sections. Here, we describe an application for AT that combines near-native tissue preservation via high-pressure freezing and freeze substitution with super-resolution light microscopy and high-resolution scanning electron microscopy (SEM) analysis on the same section. We established protocols that combine SEM with structured illumination microscopy (SIM) and direct stochastic optical reconstruction microscopy (dSTORM). We devised a method for easy, precise, and unbiased correlation of EM images and super-resolution imaging data using endogenous cellular landmarks and freely available image processing software. We demonstrate that these methods allow us to identify and label gap junctions in Caenorhabditis elegans with precision and confidence, and imaging of even smaller structures is feasible. With the emergence of connectomics, these methods will allow us to fill in the gap-acquiring the correlated ultrastructural and molecular identity of electrical synapses.
KW  - caenorhabditis elegans
KW  - localization micoscopy
KW  - fluorescent-probes
KW  - junction proteins
KW  - resolution limit
KW  - direct stochasticoptical reconstruction microscopy
KW  - structured illumination microscopy
KW  - correlative light and electron microscopy
KW  - gap junction
KW  - neural circuits
KW  - nervous-system
KW  - image data
KW  - reconstruction
KW  - innexins
KW  - super-resolution microscopy
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-187292
VL  - 3
IS  - 4
ER  - 
TY  - JOUR
A1  - Schokraie, Elham
A1  - Warnken, Uwe
A1  - Hotz-Wagenblatt, Agnes
A1  - Grohme, Markus A.
A1  - Hengherr, Steffen
A1  - Förster, Frank
A1  - Schill, Ralph O.
A1  - Frohme, Marcus
A1  - Dandekar, Thomas
A1  - Schnölzer, Martina
T1  - Comparative proteome analysis of Milnesium tardigradum in early embryonic state versus adults in active and anhydrobiotic state
JF  - PLoS One
N2  - Tardigrades have fascinated researchers for more than 300 years because of their extraordinary capability to undergo cryptobiosis and survive extreme environmental conditions. However, the survival mechanisms of tardigrades are still poorly understood mainly due to the absence of detailed knowledge about the proteome and genome of these organisms. Our study was intended to provide a basis for the functional characterization of expressed proteins in different states of tardigrades. High-throughput, high-accuracy proteomics in combination with a newly developed tardigrade specific protein database resulted in the identification of more than 3000 proteins in three different states: early embryonic state and adult animals in active and anhydrobiotic state. This comprehensive proteome resource includes protein families such as chaperones, antioxidants, ribosomal proteins, cytoskeletal proteins, transporters, protein channels, nutrient reservoirs, and developmental proteins. A comparative analysis of protein families in the different states was performed by calculating the exponentially modified protein abundance index which classifies proteins in major and minor components. This is the first step to analyzing the proteins involved in early embryonic development, and furthermore proteins which might play an important role in the transition into the anhydrobiotic state.
KW  - life-span regulation
KW  - genes
KW  - Yolk protein
KW  - water stress
KW  - expression
KW  - tolerance
KW  - richtersius coronifer
KW  - superoxide-dismutase
KW  - caenorhabditis elegans
KW  - arabidopsis thaliana
KW  - vitellogenin
Y1  - 2012
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-134447
VL  - 7
IS  - 9
ER  -