Pressure Treatment of Polytene Chromosomes and Computer Vision Techniques for High Resolution Cytogenetic Studies
Novikov, Dmitri V.
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https://hdl.handle.net/2142/86463
Description
Title
Pressure Treatment of Polytene Chromosomes and Computer Vision Techniques for High Resolution Cytogenetic Studies
Author(s)
Novikov, Dmitri V.
Issue Date
2008
Doctoral Committee Chair(s)
Belmont, Andrew S.
Department of Study
Entomology
Discipline
Entomology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Genetics
Language
eng
Abstract
The exceptional cytology provided by the interphase chromatin organization in polytene chromosomes has made Drosophila the premier model organism for analysis of chromosome structure and behavior. A major impediment to full exploitation of polytene chromosome cytology has been the difficulty in producing high quality chromosome spreads. This is particularly true for applications requiring prior formaldehyde fixation. Individual chromosome bands are seldom resolved in typical preparations, with many appearing fused. Here we describe a new procedure for preparing polytene chromosome spreads which exploits high pressure, together with other modifications, to produce high quality chromosome spreads whose light microscopy images reproducibly show comparable detail to electron microscopy images of conventionally prepared spreads. We illustrate results obtained with this procedure from typical cytological applications, including immunofluorescence and fluorescence in situ hybridization (FISH). Description of optimized downstream staining procedures for light and electron microscopy is included. Development of computer vision applications for automated recognition and structural analysis of polytene chromosomes are discussed. Prospective and collaborative studies are described.
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