Buccal micronucleus cytome assay.
The Buccal Micronucleus Cytome (BMCyt) assay is a minimally invasive method for studying DNA damage, chromosomal instability, cell death and the regenerative potential of human buccal mucosal tissue. This method is increasingly used in molecular epidemiological studies for investigating the impact of nutrition, lifestyle factors, genotoxin exposure and genotype on DNA damage, chromosome malsegregation and cell death. The biomarkers measured in this assay have been associated with increased risk of accelerated ageing, cancer and neurodegenerative diseases. This protocol describes one of the current established methods for buccal cell collection using a small-headed toothbrush, the generation of a single-cell suspension, slide preparation using cytocentrifugation, fixation and staining using Feulgen and Light Green for both bright field and fluorescence microscopic analysis. The scoring criteria for micronuclei and other nuclear anomalies are also described in detail. The protocol in its current form takes approximately 4 h to complete from the time of buccal cell collection to the generation of stained slides for microscopic analysis.
Thomas
Philip
P
CSIRO Human Nutrition, Adelaide, South Australia.
Holland
Nina
N
Bolognesi
Claudia
C
Kirsch-Volders
Micheline
M
Bonassi
Stefano
S
Zeiger
Errol
E
Knasmueller
Siegfried
S
Fenech
Michael
M
D000328:Adult; D016477:Artifacts; D016923:Cell Death; D002454:Cell Differentiation; D002455:Cell Division; D002467:Cell Nucleus / Q000502:physiology / Q000648:ultrastructure; D002843:Chromatin / Q000502:physiology; D004249:DNA Damage; D005260:Female; D006801:Humans; D007621:Karyotyping; D008297:Male; D015162:Micronucleus Tests / Q000379:methods*; D008875:Middle Aged; D009061:Mouth Mucosa / Q000166:cytology* / Q000502:physiology; D012016:Reference Values; D012038:Regeneration; D018401:Sample Size
D002843:Chromatin