The new Lamin-Chromobody® is a major breakthrough especially in High-Content Analysis (HCA) as apoptosis assays thus far rely on static endpoint assays.
The nuclear lamina is a meshwork enclosing the entire nucleus - the control center of eukaryotic cells. Its function is to hold together nuclear content and to add mechanical support to the nucleus. The nuclear lamina is a very sensitive subcellular structure. It has to maintain a certain rigidity and at the same time allow structural dynamics. Therefore, artificial incorporation of overexpressed and fluorescently labeled lamin protein mostly leads to undesired cytotoxic side effects. The new Lamin-Chromobody® binds to Lamin A/C and overcomes these unwanted side effects:
- Visualize the nuclear lamina without interfering with its cellular functions.
- Monitor the nuclear integrity and morphology during live cell microscopy as an indicator for cellular health and viability.
- Use the Lamin-Chromobody® as a biosensor for real-time assays of mitosis or apoptosis.
Confocal live cell imaging of cells going through mitosis
HeLa (left) and U2OS (right) cells stably expressing Lamin-Chromobody® fused to TagGFP. Cells going through mitosis show the typical rim like structure (in G phase), nuclear lamina depolymerization (in prometaphase) and reassembly at the end of mitosis (in telophase).
Apoptosis Assay with Lamin-Chromobody®
Time lapse imaging of untreated cells (left) and after addition of 5 µM staurosporine (STS) (right).
Quantitative evaluation with automated image acquisition and computational pattern recognition. Morphological changes of the lamina (as detected with the Lamin-Chromobody®) like fragmentation and vesicle formation were monitored in 15 minute intervals. The percentage of apoptotic cells after treatment with increasing concentrations of STS (0-5 µM) over a time of 20h is shown.