+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

In vitro micronucleus assay for the analysis of the total particulate matter in cigarette smoke comparison of flow cytometry and laser scanning cytometry with microscopy



In vitro micronucleus assay for the analysis of the total particulate matter in cigarette smoke comparison of flow cytometry and laser scanning cytometry with microscopy







The possible genotoxicity of the total particulate matter (TPM) in cigarette smoke has typically been evaluated using the in vitro micronucleus assay. In recent years, automated scoring techniques have been developed to replace the manual counting process in this assay. However, these automated scoring techniques have not been applied in routine genotoxicity assays for the analysis of TPM to improve the assay efficiency. Chinese hamster ovary (CHO) cells were treated with TPM produced from 14 types of cigarettes at five concentrations (25-200 g/ml) without exogenous metabolic activation. The three following methods were used to score the micronucleus (MN) frequency: (a) flow cytometry with SYTOX and EMA dyes, which differentially stain micronuclei and apoptotic/necrotic chromatin to enhance assay reliability; (b) laser scanning cytometry with FITC and PI dyes, which is a system that combines the analytical capabilities of flow and image cytometry; and (c) visual microcopy with Giemsa dye. The test results obtained using the three methods were compared using correlation analysis. The key findings for this set of compounds include the following: (a) both flow cytometry- and laser scanning cytometry-based methods were effective for MN identification, (b) the three scoring methods could detect dose-dependent micronucleus formation for the 14 types of TPM, and (c) the MN frequencies that were measured in the same samples by flow cytometry, laser scanning cytometry, and visual microscopy were highly correlated, and there were no significant differences (p >0.05). In conclusion, both flow cytometry and laser scanning cytometry can be used to evaluate the MN frequency induced by TPM without exogenous metabolic activation. The simpler and faster processing and the high correlation of the results make these two automatic methods appropriate tools for use in in vitro micronucleus assays for the analysis of TPM using CHO cells.

Accession: 036935390

Download citation: RISBibTeXText


Related references

In vitro micronucleus assay for the analysis of total particulate matter in cigarette smoke: comparison of flow cytometry and laser scanning cytometry with microscopy. Mutation Research 755(2): 120-125, 2013

DNA ploidy analysis by laser scanning cytometry (LSC) in colorectal cancers and comparison with flow cytometry. Cytometry 23(2): 106-109, 1996

Comparison of flow cytometry and laser scanning cytometry for the analysis of CD34+ hematopoietic stem cells. Cytometry. Part a 57(2): 100-107, 2004

Quantitative determination of Bcl-2 in FACS-sorted normal and AML progenitor cells by laser scanning cytometry Comparison with flow cytometry and Western blot analysis. Annals of Hematology 80(Suppl. 2): S32, 2001

Flow cytometry and laser scanning cytometry, a comparison of techniques. Journal of Clinical Monitoring and Computing 24(4): 251-259, 2010

Laser scanning cytometry for automation of the micronucleus assay. Mutagenesis 26(1): 153-161, 2011

Comparison of flow cytometry and laser scanning cytometry for the intracellular evaluation of adenoviral infectivity and p53 protein expression in gene therapy. Cytometry 33(3): 290-296, 1998

A comparative evaluation of single and multiparametric DNA-content analysis using flow cytometry and laser scanning cytometry. Cytometry (Suppl. 9): 124, 1998

Analysis of phytoplankton communities in lakes and artificial systems by flow cytometry and laser scanning cytometry. Cytometry Suppl. (10): 31, 2000

Automation of mouse micronucleus genotoxicity assay by laser scanning cytometry. Cytometry 44(2): 153-155, 2001

Automation of the buccal micronucleus cytome assay using laser scanning cytometry. Methods in Cell Biology 102: 321-339, 2011

Automation of the cell line based micronucleus assay by laser scanning cytometry. Toxicology Letters (Shannon) 135(Suppl 1): S40, September 15th, 2002

Flow cytometry and laser scanning confocal microscopy analysis of the receptor distribution. Nihon Yakurigaku Zasshi. Folia Pharmacologica Japonica 106(4): 255-261, 1995

Comparison of flow and laser scanning cytometry for the assay of cell proliferation in human solid tumors. Cytometry 33(3): 355-361, 1998

Automation of the cytokinesis-block micronucleus cytome assay by laser scanning cytometry and its potential application in radiation biodosimetry. Biotechniques 57(6): 309-312, 2014