Abstract

The recently developed hardware and software, in a generalized resolution overlapping fashion, now allows 3-dimensional reconstruction of subcellular structures at both the light and electron microscopic levels. It is proposed to apply these 3-dimensional techniques to the structural determination of specific Drosophila polytene bands and interbands. This study will build on the recent molecular alignment of the Notch 5'regulatory and transcribed sequences to the interband and band of polytene locus 3C7 respectively. This study will start with further molecular band/interband alignments for diverse genetic loci with a superimposition of specific proteins localized by monoclonal and affinity purified polyclonal antibodies. A detailed, 3-dimensional reconstruction at both the optical and EM levels for specific bands/interbands will proceed. Specialized software will be written to interpret and compare the complicated images. Methods will be developed for the study in 3- dimensions of specific DNa probes in situ hybridized to bands/interbands. This polytene chromosome study will be extended by a 3-dimensional reconstruction of bands/interbands under conditions of biological modification. Transcription (puff formation) and specific chromosome aberrations (position-effect varigation and transvection) are examples for structural band/interband determinations.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025101-15
Application #
3272771
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1978-04-01
Project End
1994-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
15
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143

  Publications

Shao, L; Winoto, L; Agard, D A et al. (2012) Interferometer-based structured-illumination microscopy utilizing complementary phase relationship through constructive and destructive image detection by two cameras. J Microsc 246:229-36
Matsuda, Atsushi; Shao, Lin; Boulanger, Jerome et al. (2010) Condensed mitotic chromosome structure at nanometer resolution using PALM and EGFP- histones. PLoS One 5:e12768
Kner, Peter; Winoto, Lukman; Agard, David A et al. (2010) Closed loop adaptive optics for microscopy without a wavefront sensor. Proc SPIE Int Soc Opt Eng 7570:
Kner, P; Sedat, J W; Agard, D A et al. (2010) High-resolution wide-field microscopy with adaptive optics for spherical aberration correction and motionless focusing. J Microsc 237:136-47
Chang, Henry H; Falick, Arnold M; Carlton, Peter M et al. (2008) N-terminal processing of proteins exported by malaria parasites. Mol Biochem Parasitol 160:107-15
Schermelleh, Lothar; Carlton, Peter M; Haase, Sebastian et al. (2008) Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy. Science 320:1332-6
Gustafsson, Mats G L; Shao, Lin; Carlton, Peter M et al. (2008) Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination. Biophys J 94:4957-70
Zheng, Shawn Q; Kollman, Justin M; Braunfeld, Michael B et al. (2007) Automated acquisition of electron microscopic random conical tilt sets. J Struct Biol 157:148-55
Xu, Jingsong; Van Keymeulen, Alexandra; Wakida, Nicole M et al. (2007) Polarity reveals intrinsic cell chirality. Proc Natl Acad Sci U S A 104:9296-300
Krug, Roland; Carballido-Gamio, Julio; Burghardt, Andrew J et al. (2007) Wavelet-based characterization of vertebral trabecular bone structure from magnetic resonance images at 3 T compared with micro-computed tomographic measurements. Magn Reson Imaging 25:392-8

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