Pressure Treatment of Polytene Chromosomes and Computer Vision Techniques for High Resolution Cytogenetic Studies

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.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Entomology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Novikov, Dmitri V.

Contributors dc:contributor

• Belmont, Andrew S.

• Biology, Genetics

Language dc:language

eng

Identifier

(MiAaPQ)AAI3314860

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/86463