• NEWS

***********************

Publication 2019 Nov:
Direct, gabapentin-
insensitive interaction
of a soluble form of
the calcium channel subunit
α2δ-1 with thrombospondin-4
https://www.ncbi.nlm
.nih.gov/pubmed/31700036

***********************

***********************

Publication 2019 Oct:
Dual Inhibition of
GLUT1 and the ATR/CHK1
Kinase Axis Displays
Synergistic Cytotoxicity
in KRAS-Mutant Cancer Cells.
https://www.ncbi.nlm
.nih.gov/pubmed/31405847

***********************

Publication 2019
High-Content Imaging
of Immunofluorescently
Labeled TRPV1-Positive
Sensory Neurons.
https://www.ncbi.nlm.
nih.gov/pubmed/31028677

Publication 2019 Jan: UBQLN4 Represses Homologous Recombination and Is Overexpressed in Aggressive Tumors. https://www.ncbi.nlm.nih
.gov/pubmed/30612738

***********************

Publication 2018 Oct: The Cdkn1aSUPER Mouse as a Tool to Study p53-Mediated Tumor Suppression. https://www.ncbi.nlm.
nih.gov/pubmed/30355482

***********************

Publication 2018 June:
Evaluation of small
intestinal damage in
a rat model of 6 Minutes
cardiac arrest.
https://www.ncbi.nlm.nih
.gov/pubmed/29866034

***********************

Publication 2018 May:
A Hierarchical, Data-Driven
Approach to Modeling
Single-Cell Populations
Predicts Latent Causes of
Cell-To-Cell Variability.
https://www.ncbi.nlm.
snih.gov/pubmed/29730254

***********************

Publication 2018 April: PKA-RII subunit phosphorylation precedes activation by cAMP and regulates activity termination.
https://www.ncbi.nlm.
nih.gov/pubmed/
29615473

***********************

 

 

 

 

APPROACHES

Central to our work is to get as quantitative data as possible. At the core of our work is an automated “High Content Screening” microscopy approach.
In short: large numbers of pictures of e.g. cell cultures are taken. Then in a second step these digitalized data are quantified via automated object identification algorithms. Thereby not only one but many parameters can be read out such as cell numbers, cell size, morphological characteristica, absolute and relative position to each other, marker intensity, subcellular localization, …. This allows us to reliably quantify in an exactly defined manner any aspect of the images. This technique can be applied not only to pain research but also to any other field, which results in images. Examples of what we have been quantifying so far shown below. Please feel free to contact us, if you would love to collaborate with us on, well, any kind of quantification, be it simple or complex.

Workflow to quantify cellular signaling in subgroups of sensory neurons.
Figure 1: Workflow to quantify cellular signaling in subgroups of sensory neurons.
Dorsal root ganglia (DRGs) are collected, enzymatically digested, and cultured overnight. Subsequent to stimulation with e.g. inflammatory mediators, cells are either fixed and stained with phospho-specific kinase antibodies, imaged alive (Ca2+ imaging), or combinations of both. Image acquisition is performed with an automated Cellomics ArrayScan microscope. Images are analyzed by automated image analysis resulting in quantitative single cell data. (Images by Jörg Isensee)
Quantification of marker protein expression sections of dorsal root ganglia.
Figure 2: Quantification of marker protein expression sections of dorsal root ganglia.
Confocal images of lumbar DRG sections triple-stained for UCHL1, RIIβ, and IB4 (10 µM thick, scale bar = 50 µM). The image of the neuronal marker UCHL1 was used to generate an image mask allowing the quantification of signal intensities in other fluorescence channels. (Images by Jörg Isensee)
Figure 3: Analysis of sciatic nerve sections.
Figure 3: Analysis of sciatic nerve sections. Phase contrast images of sciatic nerve sections were analyzed to quantify the diameter and number of fibers. (Image by Ilja Bobylev / Jörg Isensee)
High content microscopy approach to investigate stress granule formation in cell lines.
Figure 4: High content microscopy approach to investigate stress granule formation in cell lines.
(Images by Christian Kähler / Jörg Isensee)
Quantification of CREB phosphorylation within neuronal nuclei
Figure 5: Quantification of CREB phosphorylation within neuronal nuclei.
Immunostaining of phospho-CREB in DRG neurons stimulated for 15 min with forskolin (10 µM) or solvent control (0.1% DMSO) (Scale bar = 20 μm). Image analysis allows the specific analysis of neuronal nuclei. (Images by Jörg Isensee)
Neuronal and glial signaling
Figure 6: Neuronal and glial signaling: We use the high content screening microscopy (HCS) in combination with confocal microscopy to quantify the signaling pathways activated upon stimulation in DRG cultures. In particular we are interested in qualitative and quantitative differences in signaling in sensory neurons and the different associated glial cell types in the ganglia. The goal is to connect these signaling pathways with functional outcomes in both cell types.