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Dr Brian T Bennett 4Pi Microscopy
4 Pi Microscopy the Human Nuclei DNA Damage Follwing Ionizing Radiation

Dr. Brian T. Bennett, Assistant Research Professor, University of Utah  completes the assembly of a super-resolution FPALM, Biplane microscope.  Biplane microscopy provides three-dimensional sub-100nm resolution (~30X30X75nm) of thick samples without scanning. The method employs the dual plane detection system of Biplane microscopes to achieve super-resolution in the z-axis.  Biplane methodology is combined with fluorescent photoactivatable localization microscopy (FPALM) to enable 3D sub-diffraction resolution without scanning. 

 

Shown here is a dual-plane single molecule detection of the mitochondria 

Dr Brian T Bennett 4Pi Microscopy
4 Pi Microscopy the Human Nuclei DNA Damage Follwing Ionizing Radiation

Three-dimensional video representation of a 4Pi microscopy data set showing nuclear distribution of H2AX and g-H2AX at 15 min after exposure to ionizing radiation.   Video shows a single nuclei within a human immortalized cell line.  H2AX is in green and g-H2AX is in red. The grid has a 1-mm raster.

Bennett et al., Proceedings of the National Academy of Science

Shown here is the first 4Pi Super-Resolution image captured using the Leica Commercial 4Pi System of a biological sample.  HeLa immortalized cell line in division.  Cell preparation, staining of  α-tubulin, and fixation by Dr. Brian T. Bennett.  Image by Dr. Brian T. Bennett and Dr. Tanjef Zellas, Leica Microsystems, 2005. 

Dr Brian T Bennett 4Pi Microscopy
4 Pi Microscopy the Human Nuclei DNA Damage Follwing Ionizing Radiation

 Three-dimensional video representation of a 4Pi microscopy data set showing nuclear distribution of H2AX and g-H2AX at 15 min after exposure of cells to ionizing radiation.   H2AX is in green and g-H2AX is in red. The grid has a 1-m m raster and axes are as indicated in Fig. 1.

Comparison of confocal vs. 4Pi microscopy for endogenous human histone H2AX. (a and b) The data shown represent the H2AX staining in a HeLa cell nucleus as seen with confocal and 4Pi microscopy, respectively, in a maximum projection of 3D data sets. (c and d) The same single xz section taken from the 3D data sets of a and b, respectively. The dotted ellipses mark areas where the resolution enhancement can be seen best. (c Inset and d Inset) The PSFs of confocal and 4Pi microscopy at the same scale as a–d for comparison.

© Copyright 2003 -2018 Dr. Brian T Bennett PLLC, All images, publications, articles etc are the work of Dr. Brian T. Bennett, and collaborators where noted, and are open source, except where password protected.