Movie Cell-Nucleus_stack.avi
MovieCell-Nucleus_stack.avi
Confocal Laser Scanning Microscpy image stack of a Chironomus
tentans salivary gland cell nucleus. The fluorescence was due to
the protein hrp36 conjugated to red-fluorescent AlexaFluor647,
which was microinjected into the cytoplasm. After transport into
the nucleus, hpr36 was incorporated into nascent RNA-particles,
such that active transcription sites were marked. The brighter oval
region in the image center is the cell nucleus. The polytene
chromosomes showed their characteristic banding pattern. The images
were aquired 15 min after after microinjection.Complete cell in
fluorescence color-coded by the look-up-table red hot of ImageJ
Each image comprises 772x512 pixels, a total volume of 185 m x 29 m
x 46 m was imaged using a C-Apochromat 40X (NA 1.2 , water
immersion) objective lens. The pinhole was set to 1.2 Airy units.
Excitation 633nm (5% output); main dichromat UV/488/543/633;
emission band pass filter LP650.
Data courtesy of Dr. Jan Peter Siebrasse and Dr. Ulrich
Kubitscheck, Institute of Physical and Theoretical Chemistry,
Rheinische Friedrich-Wilhelms-Universtitt Bonn, 2011 (For further
information on the biological contents, please refer to:Siebrasse.
J.P., T.Kaminski and U.Kubitscheck (2012) Nuclear export of single
native mRNA molecules observed by light sheet fluorescence
microscopy. Proc Natl Acad Sci USA 109: 9426-31)
MovieCell_Nucleus_rotation.avi
The above image stack was processed and optimized for a 3D
rotation presentation. All data processing was done using ImageJ by
Wayne Rasband, NIH (see http://rsb.info.nih.gov/ij/).Data courtesy
of Dr. Jan Peter Siebrasse and Dr. Ulrich Kubitscheck, Institute of
Physical and Theoretical Chemistry, Rheinische
Friedrich-Wilhelms-Universtitt Bonn, 2011 (For further information
on the biological contents, please refer to:Siebrasse. J.P.,
T.Kaminski and U.Kubitscheck. 2012. Nuclear export of single native
mRNA molecules observed by light sheet fluorescence microscopy.
Proc Natl Acad Sci USA 109: 9426-31)
MovieConfocal_PSF.mp4
Confocal Point-Spread-FunctionCalculated isointensity surfaces
of the three-dimensional confocal point-spread-function are shown
for a 0.7 NA objective with a 1 AU pinhole. The integrated
intensity of the confocal psf is projected onto the individual axes
and colored according to the color bar on the right. The
isointensity surface shows all values of the psf having the same
intensity relative to the maximum of the psf and is plotted using
the same color table. We gratefully acknowledge Nicolai Hartmann,
Department of Chemistry and Center for Nanoscience (CeNS),
Ludwig-Maximilians-University Munich, for producing this
movie.MoviePSF_Pinhole.mp4
Confocal Point Spread Function as a function of Pinhole sizeThis
video shows how the confocal point spread functions (psf) changes
as the size of the pinhole is varied. The excitation psf without
pinhole is shown in the upper left panel and the total psf is shown
in the right upper panel. The fraction of light that is transmitted
through the pinhole is shown on the graph in the lower panel with
the green cursor marking the current pinhole size for the
calculated psf. The smaller the pinhole size yields a higher
overall resolution but at a significant loss in detection yield.
The lower panel shows that a pinhole with the size of approximately
0.8 1 AU is the optimal compromise between resolution and detection
yield. We thank Achim Hartschuh, Physical Chemistry, Department of
Chemistry and Center for Nanoscience (CeNS),
Ludwig-Maximilians-University of Munich, Germany, for producing
this movie.MovieConfocal_GFP_Cell.mov
Confocal Image of a Huh7 cell with eGFP-a-tubulin. This movie
shows a three-dimensional reconstruction of a z-stack of confocal
images from a Huh7 cell, where a-tubulin has been tagged with eGFP.
The microtubules are clearly visible, demonstrating the capability
of confocal microscopy for three-dimensional imaging. The diameter
of the cell is approximately 60 m. Dr. Ralf Bausinger, Department
of Chemistry, University of Konstanz, Germany, is gratefully
acknowledged for collecting and processing the data and for making
this video available to us.MovieSingle_Molecules
Single DiI molecules diffusing in a fluid glass-supported lipid
bilayer. The bilayer was prepared by vesicle-fusion from the lipid
dioleyl-phosphatidylcholine (DOPC) with low amounts of the
lipohilic dye DiI (molar fraction of ~10-8). The detail has a size
of 30 x 30 m2, recording rate was 20 frames per second, the movie
plays at real-time. Data Courtesy by E. Klotzsch and G.J.Schtz,
Institute of Applied Physics, Vienna University of Technology,
Austria