Abstract：
BACKGROUND:Assessment of resistance to drug moieties in tissue culture is complicated by limited sample, clonal selection and alteration of cycling fraction and cycle duration in clonally mixed lesions. DNA damage assessment by single cell gel electrophoresis (SCGE) of excised DNA is limited by non-linear analysis in fluorescent light microscopy. Confocal Laser Scanning Microscopy (CLSM) with high N.A. magnification allows for quantitation of total excised DNA fragment size distribution but is still limited by the large volume required for labour intensive SCGE, precluding multi-exposure clinical testing. AIMS:To optimise sample requirement for SCGE and CLS. METHODS:Standard slide mounted bed gels were punched with multiple coded 6 mm wells and filled with suspensions of cells subjected to drug/concentration variations. After SCGE, 30 microns frozen sections were prepared of each well and mounted in ethidium bromide solution on multi-well hydrophilic slides to allow for short working distance of high resolution CLSM in a Zeiss Axiovert L410 SM. Testing for feasibility, reproducibility and consistency used both cultured standard leukaemic cell lines, normal human control marrow and clinical samples. RESULTS AND CONCLUSION:Multiple well SCGE followed by frozen section, high resolution CLSM allows for rapid analysis of high numbers of multiple drug exposure permutations clinically required.
journal_name
Adv Exp Med Bioljournal_title
Advances in experimental medicine and biologyauthors
Lannon CL,Ball LM,Pyesmany AF,Yhap M,Langley R,van Velzen Ddoi
10.1007/978-1-4615-4811-9_57subject
Has Abstractpub_date
1999-01-01 00:00:00pages
527-35eissn
0065-2598issn
2214-8019journal_volume
457pub_type
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