Role of water in chromosome spreading and swelling induced by acetic acid treatment: a FTIR spectroscopy study

Submitted: 8 October 2013
Accepted: 30 December 2013
Published: 11 February 2014
Abstract Views: 2984
PDF: 837
Supplementary: 229
HTML: 711
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Authors

The so called chromosome preparation is a procedure consisting of three strictly connected stages that enables to obtain chromosomes of quality suitable for cytogenetic analysis. Interestingly, experimental evidence strongly suggested that chromosome spreading and swelling (key processes that allow their counting and detailed structural analysis) are induced in the last fixative-evaporation stage by the interaction, mediated by acetic acid, between water from the environmental humidity, and the cytoplasmic matrix and the chromatin. However, since a considerable variation in the quality of chromosome preparations is observed, strongly depending on the environmental conditions in which the procedure takes place, a better comprehension of the mechanisms underlying chromosome preparation is required. To this aim, here we analysed intact lymphocytes before and at each stage of the chromosome preparation protocol by Fourier transform infrared (FTIR) spectroscopy, a technique widely used for the study not only of isolated biomolecules, but also of complex biological systems, such as whole cells. Interestingly, we found that the chromosome preparation protocol induces significant structural changes of cell proteins and DNA, in particular due to the interaction with acetic acid. Moreover, noteworthy, through the monitoring of changes in the water combination band between 2300 and 1800 cm–1, we provided evidence at molecular level of the crucial role of the bound water to the cytoplasmic matrix and to the chromatin in determining the chromosome spreading and swelling. Our FTIR results, therefore, underline the need to perform the last fixative-evaporation stage in standardized and optimized temperature and relative humidity conditions, thus providing chromosomes of high quality for the cytogenetic analysis that would lead in this way to more reliable results.

Dimensions

Altmetric

PlumX Metrics

Downloads

Download data is not yet available.

Citations

V. Meraviglia, Centro Cardiologico Monzino
Vascular Biology and Regenerative Medicine Lab
M. Baccarin, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico
Laboratory of Medical Genetics
G. Terzoli, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico
Laboratory of Medical Genetics

How to Cite

Ami, D., Di Segni, M., Forcella, M., Meraviglia, V., Baccarin, M., Doglia, S., & Terzoli, G. (2014). Role of water in chromosome spreading and swelling induced by acetic acid treatment: a FTIR spectroscopy study. European Journal of Histochemistry, 58(1). https://doi.org/10.4081/ejh.2014.2330

Similar Articles

1 2 3 4 5 6 7 8 9 10 > >> 

You may also start an advanced similarity search for this article.

Publication Facts

Metric
This article
Other articles
Peer reviewers 
2
2.4

Reviewer profiles  N/A

Author statements

Author statements
This article
Other articles
Data availability 
N/A
16%
External funding 
N/A
32%
Competing interests 
N/A
11%
Metric
This journal
Other journals
Articles accepted 
57%
33%
Days to publication 
125
145

Indexed in

Editor & editorial board
profiles
Academic society 
N/A