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Topics: 1.Preparation 2.Aging 3.Denaturing 4.Methods 5.Table | Fig1 | Fig2 |
Table 1. Result comparisons between the standard and the proposed spreading protocols.
|
Periph blood |
BM1 |
BM1 |
BM1 |
BM2 |
BM2 |
|
|
Standard laboratory protocol |
||||||
|
No. metaphases |
57 |
50 |
53 |
42 |
45 |
38 |
|
Avg diam. (um) |
32.83 |
17.76 |
20.56 |
18.48 |
16.97 |
16.89 |
|
Stand. Dev. |
6.55 |
3.85 |
7.28 |
3.58 |
3.89 |
2.7 |
|
No. crossov |
242 |
NA |
NA |
NA |
NA |
NA |
|
Cross/metaph. |
4.3 |
NA |
NA |
NA |
NA |
NA |
|
Proposed spreading protocol |
||||||
|
No metaphases |
85 |
58 |
45 |
47 |
35 |
47 |
|
Avg diameter |
39.41 |
22.31 |
24.21 |
22.95 |
22.26 |
21.37 |
|
Stand. Dev. |
4.5 |
5.64 |
8.04 |
6.49 |
4.06 |
4.88 |
|
No crossov |
115 |
NA |
NA |
NA |
NA |
NA |
|
Cross/metaph. |
1.4 |
NA |
NA |
NA |
NA |
NA |
Table 1 legend: Cells from a peripheral blood culture and two or three separate flasks from two bone marrow cultures were subjected to both the standard and the proposed slide preparation technique. Each slide was scanned from left to right and top to bottom using the 20x objective. All metaphases found (all fields containing at least one metaphase) were captured, to a final number of 36 microscopic fields on each slide (45 fields for the peripheral blood slide obtained with the proposed protocol). The diameter of every captured metaphase was measured (in microns). The number of crossovers for the longer, peripheral blood culture chromosomes was also recorded. As chromosomes from bone marrow cultures were usually short, the number of crossovers was reduced and not so informative, thus it was not included in the study. No. metaphases indicates the number of metaphases captured and measured. Avg. diam. indicates the arithmetic mean of all diameters measured. Stand. Dev. is the standard deviation of the values measured. No. crossov. are the number of crossovers found in all metaphases captured. Crossov/metaph. indicates the average number of crossovers per metaphase. BM1 and BM2 are the two bone marrow cultures analyzed. GCT or EB are the different culture flasks used for the same BM culture