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Imaris view of a
low-pass filtered 3D image data stack displaying two GFP-tag, which indicate
the position of the spindle pole body (green) and the centromere of chromosome
IV (red) in interphase (G1).
The trajectory has
been measured by LCCB's own ChromDyn software package (authors: Jonas
Dorn and Khuloud Jaqaman, LCCB) which calls Imaris for complex visualization
and data representation via a prototype version of the new ImarisXT interface.
This interface allows
us to effortlessly merge application-specific image analysis with the
powerful and general purpose 3D visualization of Imaris. Special features
in ChromDyn are:
- Super-resolution
capability, i.e. the tracker can follow two or more tags at distances
up to three times below the diffraction limit of light microscopy.
- Low signal to noise
detection, i.e. the tracker can cope with tag signals that are not more
than 2 - 3 times brighter than the noise of raw data;
- Mutant phenotyping,
i.e. the package includes statistical evaluation of tag trajectories
in the framework of a physical model describing the regulated dynamics
of the microtubules linking the two tags (or more, when analyzing the
arrangement of a bipolar spindle in metaphase) in order to sensitively
screen mutants for protein function.
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The Laboratory
for Computational Cell Biololgy (LCCB) is part of the Center for Integrative
Molecular Biosciences (CIMBio) at the Scripps Research Institute (TSRI).
CIMBio was founded in 2002 to foster multidisciplinary studies of molecular
machines, with the aim of determining their structure, their mechanism
of action, and their behavior in the context of living cells and whole
organisms in normal physiology and disease. The LCCB contributes to this
effort by developing quantitative light microscopy and numerical models
of cytoskeleton mechanics to define the relationship between the dynamics
and control of cytoskeletal protein assemblies and cellular responses
across space and time scales.
Specifically, we investigate:
- The actin cytoskeleton
as a multi-functional, complex molecular system and how its dynamics
mediate processes such as cell migration, morphogenesis, phagocytosis,
endocytosis, virus infection, and intra-cellular transport. Our ultimate
goal is to establish an integrated quantitative model for how regulatory
molecules change the biochemical and mechanical properties of the actin
cytoskeleton at the ultra-structural level in order to support these
diverse functions, and to test the model in living cells using quantitative
Fluorescent Speckle Microscopy.
- How microtubule
dynamics mediate chromosome segregation during mitosis. We are developing
super-resolution microscopy and structural models of the mitotic spindle
apparatus in yeast to exploit the power of yeast genetics in performing
functional screens for regulatory proteins of microtubule dynamics and
force generation. In parallel, we work on extending Fluorescent Speckle
Microscopy to 3D with the aim to study these processes in higher organisms.
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