ChromaWizard: An open source image analysis software for multicolor fluorescence in situ hybridization analysis

Abstract

Multicolor image analysis finds its applications in a broad range of biological studies. Specifically, multiplex fluorescence in situ hybridization (M‐FISH) for chromosome painting facilitates the analysis of individual chromosomes in complex metaphase spreads and is widely used to detect both numerical and structural aberrations. While this is well established for human and mouse karyotypes, for which species sophisticated software and analysis tools are available, other organisms and species are less well served. Commercially available software is proprietary and not easily adaptable to other karyotypes. Therefore, a publically available open source software that combines flexibility and customizable functionalities is needed. Here we present such a tool called “ChromaWizard” which is based on popular scientific image analysis libraries (OpenCV, scikit‐image, and NumPy). We demonstrate its functionality on the example of primary Chinese hamster (Cricetulus griseus) fibroblasts metaphase spreads and on Chinese Hamster Ovary cell lines known for the large number of chromosomal rearrangements. The application can be easily adapted to any kind of available labeling kits and is https://dict.leo.org/englisch-deutsch/independent of the used organism and instrumentation. It allows direct inspection of the original hybridization signals and enables either manual or automatic assignment of colors, making it a functional and versatile tool that can be used also for other multicolor applications.

Chromosomal instability drives convergent and divergent evolution toward advantageous inherited traits in mammalian CHO bioproduction lineages

“…We next reasoned that numerical chromosome heterogeneity was insufficient to determine genomic stability of these cell lines. Chromosome number can appear to be constant while rearrangements and other structural changes occur ( Auer et al., 2018 ; Brinkrolf et al., 2013 ; Vcelar et al., 2018 ). Other studies have noted that the configuration and copy number of rearranged chromosomes are associated with productivity in CHO ( Yamano et al., 2015 ).…”

A reliable technique for karyotyping mouse oocytes prepared by a gradual fixation/air-drying method followed by multicolour FISH

“…The images of five fluorescent colours (aqua, green, orange, red and near-infrared) and DAPI of the chromosome spreads were acquired using a fluorescence microscope (BX51; Olympus, Tokyo, Japan) with a high-sensitivity digital camera (α7 s; Sony, Tokyo, Japan). All images were imported into ChromaWizard software ( Auer et al, 2018 ), and karyotype analysis was performed in the ChromaWizard software.…”

Birth of mice from meiotically arrested spermatocytes following biparental meiosis in halved oocytes

“…Fluorescence images were captured using a high‐sensitive digital camera (α7s, SONY). The images were imported into the ChromaWizard software (Auer et al , 2018) to assign fluorescence colors to each chromosome. Based on these fluorescence colors, the chromosome numbering was determined.…”