Albumin enhanced morphometric image analysis in CLL.
BACKGROUND
The heterogeneity of lymphocytes from patients with chronic lymphocytic leukemia (CLL) and blood film artifacts make morphologic subclassification of this disease difficult.


METHODS
We reviewed paired blood films prepared from ethylene-diamine-tetraacetic acid (ETDA) samples with and without bovine serum albumin (BSA) from 82 CLL patients. Group 1 adhered to NCCLS specifications for the preparations of EDTA blood films. Group 2 consisted of blood films containing EDTA and a 1:12 dilution of 22% BSA. Eight patients were selected for digital photomicroscopy and statistical analysis. Approximately 100 lymphocytes from each slide were digitally captured.


RESULTS
The mean cell area +/- standard error was 127.8 microm(2) +/- 1.42 for (n = 793) for group 1 versus 100.7 microm(2) +/- 1.39 (n = 831) for group 2. The nuclear area was 88.9 microm(2) +/- 0.85 for group 1 versus 76.4 microm(2) +/- 0.83 for group 2. For the nuclear transmittance, the values were 97.6 +/- 0.85 for group 1 and 104.1 +/- 0.83 for group 2. The nuclear:cytoplasmic ratios were 0.71 +/- 0.003 for group 1 and 0.78 +/- 0.003 for group 2. All differences were statistically significant (P < 0.001).


CONCLUSIONS
BSA addition results in the reduction of atypical lymphocytes and a decrease in smudge cells. BSA also decreases the lymphocyte area and nuclear area, whereas nuclear transmittance and nuclear:cytoplasmic ratio are increased. A standardized method of slide preparation would allow accurate interlaboratory comparison. The use of BSA may permit better implementation of the blood film-based subclassification of CLL and lead to a better correlation of morphology with cytogenetics and immunophenotyping. Published 2003 Wiley-Liss, Inc.
Lunning
Matthew A
MA
Flow and Image Cytometry Section, Laboratory Stem Cell Biology, Division of Cell and Gene Therapies, Center for Biologics Research and Evaluation, Food and Drug Administration, Bethesda, Maryland 20892, USA.
Zenger
Vincent E
VE
Dreyfuss
Ricardo
R
Stetler-Stevenson
Maryalice
M
Rick
Margaret E
ME
White
Therese A
TA
Wilson
Wyndham H
WH
Marti
Gerald E
GE
D001800:Blood Specimen Collection / Q000379:methods*; D004492:Edetic Acid; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000097:blood*; D012044:Regression Analysis; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000378:metabolism*; D012710:Serum Albumin, Bovine
D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D004492:Edetic Acid