Interactive computer-aided assignment of multiple probes to cytogenetic bands by simultaneous dual color fluorescence in situ hybridization and DAPI banding

Abstract

A macro function was developed to run in conjunction with the popular image analysis package NIH Image, to allow simultaneous determination of mapping positions of one or two separate probes with respect to cytogenetic bands by dual color fluorescence in situ hybridization (FISH) and DAPI banding, and by determination of their fractional distance from pter (FLpter). In order to allow maximal flexibility, a user‐defined line along the chromosome is used for measurements. Algorithms were developed to detect the ends of the chromosome and the cytogenetic bands. Results of the analysis are presented in graphical form, comprising a display of the DAPI intensity along the chromosome, the positions of the probe(s), the locations of bands as determined by analysis of the second derivative of the DAPI intensity profile, and a standard ideogram of the chromosome for comparison. The approach was validated and compared to visual assignment of probes to DAPI bands using the cosmid clone PYGM, which has been previously mapped to chromosome 11q13, and has been used as a landmark for mapping of other probes (3). © 1996 Wiley‐Liss, Inc.

Cloning a novel member of the human interferon-inducible gene family associated with control of tumorigenicity in a model of human melanoma

“…Map positions (cytogenetic band and Flpter measurements) were obtained for each hybridization signal in 20 metaphase chromosomes using Band Map 2.2 (Burde et al, 1996) in NIH Image.…”

Selection and application of a rainbow trout toxicity testing procedure for screening Sacramento River Watershed, California samples

“…In addition, even with 10 days of development, the embryos lacked sufficient tissue and distinct morphological features to evaluate using either low or medium magnification with or without image capture. Burde et al (1996) suggested that fluorescence imaging with protein tags can provide insight into normal versus abnormal development. Exploratory video imaging experiments in our laboratory did not provide the differential necessary to detect low-level stress to developing embryos.…”

Assessing the applicability of a modified replication banding protocol for the analysis of radiation-induced chromosomal aberrations in cultured human lymphocytes

“…The precise localization of structural chromosomal alterations provides a significant indicator to analyze genomic imbalances induced by ionizing radiation. To localize rearranged sites revealed by FISH, counterstaining with 4′,6-diamidino-2-phenylindole (DAPI) is mostly carried out to generate a banding pattern as a chromosomal landmark (Heng and Tsui 1993, Knapp et al 1995, Burde et al 1996, Barros e Silva and Guerra 2010). DAPI has an affinity to chromosomal regions with high contents of adenine and thymine residues (AT-rich regions) that correspond to late-replicating bands (Costantini and Bernardi 2008).…”