bluuebunny/crossref_metadata_2025_split huggingface dataset:
Leukemia‐related morphological features in blast cells
Authors: Aus H. M. , Harms H. , Haucke M. , Beritova J. , ter Meulen V. , Gunzer U. , Baumann I. , Abmayr W. 
Published: August 2005

AbstractThis paper investigates the use of image‐processing methods to detect leukemia‐related morphological differences in mononuclear blast cells. Routinely prepared Pappenheim‐stained blood smears were scanned in a high‐resolution color TV‐microscope system. Eleven blast‐cell classes (OMSBC, T‐ALL, OMS, ALL, LBL, IBL, AUL, AML, AMOL, AMMOL, and CML) were analyzed with the nonparametric statistical software program „Classification and Regression Trees”︁ (CART). This paper documents the initial statistical evaluation of 62 leukemia‐related morphological features that directly measure and analyze the cell‐related quantifiable differences occurring in the various blast cells. The 62 cell image features include both common cytophotometric features, and new texture and color features developed for this project. This study found that each leukemia specimen contains a dominant class of blasts that correlates with the specific leukemia, plus a distribution of blasts from related diseases. The present data suggest the existence of a distribution fingerprint pattern for each leukemia.
Can peripheral T‐Cell lymphomas be morphologically subclassified? A morphometric approach to 21 cases
Authors: Sigaux François , Flandrin Georges , Valensi Françoise , Gaulard Philippe , Castaigne Sylvie , Daniel Marie Therese , Harousseau Jean Luc 
Published: August 2005

AbstractMorphometric analysis of nuclear sizes and shapes was carried out on semithin sections of lymph node for 21 patients suffering from non‐Hodgkin's peripheral T‐cell malignant lymphoma (ML) (excluding mycosis and Sézary syndrome). Twenty cases of B‐cell ML and three cases of Sézary syndrome with massive lymph node infiltration were also studied as references. Wright and Isaacson's recent proposals were applied to classify the peripheral T‐cell MLs into monomorphic medium‐cell ML (eight cases), pleomorphic ML (nine cases), and monomorphic large‐cell ML (four cases). These three classes were readily distinguishable by morphometric analysis of nuclear sizes. Nuclear areas and their coefficients of variation were higher in pleomorphic MLs than in monomorphic medium‐cell MLs (p < 0.01). Large‐cell monomorphic MLs were set apart by the histograms of their nuclear sizes. The mitoses were evaluated on histological sections and found to be more numerous in pleomorphic ML than in monomorphic medium‐cell ML (p < 0.05). Nuclear irregularity in the 21 cases of peripheral T‐cell ML was lower than in Sézary cells. Morphometry clearly demonstrates the morphological distinctiveness of the subclasses of peripheral T‐cell ML. Their biological significance has yet to be determined.
Binding, ingestion, and growth of Chlamydia trachomatis (L2 serovar) analyzed by flow cytometry
Authors: Levitt Daniel , Zable Brian , Bard Jonathan 
Published: August 2005

AbstractWe have developed a method for quantitatively assessing binding, ingestion, and growth of Chlamydia trachomatis (L2 serovar) in several mammalian cell lines using fluorescence staining and flow cytometry. Cells were incubated with chlamydia at 4°C to monitor binding; ingestion was determined by raising the temperature to 37°C for 1–4 h and removing extracellular bacteria with pronase. Growth of bacteria was measured by assessing brightly stained intracellular inclusions. Fixation with methanol prior to fluorescent staining provided the most intense specific staining with minimal background, as well as preserving cell morphology. Our data reveal relatively slow ingestion of L2 by McCoy fibroblasts (maximum ingestion by 4 h) and a sizeable population of McCoy cells (30–40% of total cells) that ingest L2 but do not permit its growth under certain infectious conditions. It was possible to correlate specific histogram patterns on the flow cytometer with fluorescent microscope observations. This system provides a means of analyzing quantitative interactions between chlamydia and individual host cells.
Flow cytometric estimation of cell cycle parameters using a monoclonal antibody to bromodeoxyuridine
Authors: Sasaki Kohsuke , Murakami Tomoyuki , Ogino Tetsuro , Takahashi Manabu , Kawasaki Shoji 
Published: August 2005

AbstractAn estimation of cell kinetic parameters was made by simultaneous flow cytometric measurements of DNA and bromodeoxyuridine (BrdUrd) contents of cells. The procedure described in this paper involves the incorporation of BrdUrd by S phase cells, labeling the BrdUrd with an indirect immunofluoresent technique using a monoclonal anti‐BrdUrd antibody, and staining DNA with propidium iodide (PI). The amount of incorporated BrdUrd in HeLa cells was proportional to that of synthesized DNA through S phase. For all cell lines examined, the pattern of BrdUrd incorporation was essentially the same and the rate of DNA synthesis during S phase was not constant. The bivariate BrdUrd/DNA distributions showed a horse‐shoe pattern, maximum in the mid S phase and minimum in the early and late S phases. Furthermore, the durations of cell cycle (Tc) and S phase (Ts) were estimated from a FLSm (fraction of labeled cells in mid S phase) curve that was generated by plotting the percentage of BrdUrd pulse‐labeled cells in a narrow window defined in the mid S phase of the DNA histogram. The values of these parameters in NIH 3T3, HeLa S3, and HL‐60 cells were in good accordance with the reported data. This FCM method using the monoclonal anti‐BrdUrd antibody allows rapid determination of both cell cycle compartments and also Ts and Tc without the use of radioactive DNA precursors.
Construction, analysis, and application to 46,XY gonadal dysgenesis of a recombinant phage DNA library from flow‐sorted human Y chromosomes
Authors: Müller Ulrich , Lalande Marc , Disteche Christine M. , Latt Samuel A. 
Published: August 2005

AbstractThe analysis of a recombinant human Y‐enriched Hind III total digest phage library prepared from the DNA of flow sorted human Y chromosomes is described. Out of 43 phage inserts from the library thus far mapped, 25 revealed hybridization with Y chromosomal DNA. These inserts may be divided into five groups according to their degree of Y specific hybridization: inserts that hybridize with 1) one single copy or slightly repeated Y‐specific DNA sequence, 2) Y‐specific repeated sequences of various restriction fragment lengths, 3) Y‐chromosomal DNA sequence(s) shared by a sequence on the X and/or on autosomes, 4) Y‐specific DNA sequences in addition to multiple X and/or autosomal sequences, or 5) Y‐specific repeated DNA in addition to multiple X and/or autosomal sequences. Application of probes from this library for diagnostic purposes is shown in two 46, XY patients with gonadal dysgenesis and small deletions of the Y short arm.
A method to average immunofluorescent histograms
Authors: Marti G. E. , Schuette W. , Magruder L. , Gralnick H. R. 
Published: August 2005

AbstractSingle parameter gated flow cytometric fluorescent histograms were obtained on normal donor blood mononuclear cells using several commonly available lymphocyte surface markers. A computer method was developed to average single parameter flow cytometric immunofluorescent histograms. The averaged histograms provide a means of pattern recognition for normal lymphocytes and will aid in the clinical evaluation of lymphocytosis. Averaged histograms may also serve as standards for more advanced analysis.
Detection of acid‐β‐galactosidase activity in viable human fibroblasts by flow cytometry
Authors: Jongkind J. F. , Verkerk A. , Sernetz M. 
Published: August 2005

AbstractThe fluorogenic substrate fluorescein‐di‐β‐D‐galactopyranoside was used to detect acid β‐galactosidase in intact cultured human fibroblasts. The accumulation of intracellular fluorescein, as measured by flow cytophotometry was linear with the incubation time in three control strains. The two fibroblast strains from patients with acid β‐galactosidase deficiency did not show an accumulation of intracellular fluorescence. Within one control cell population there was a positive correlation between the amount of accumulated intracellular fluorescein fluorescence and the specific acid β‐galactosidase activity as measured biochemically on sorted cells from different zones of the fluorescence distribution. No correlation was found between the specific acid β‐galactosidase activity and the fluorescein fluorescence of three different control cell strains.
Characterization of chromatin distribution in cell nuclei
Authors: Young Ian T. , Verbeek P. W. , Mayall Brian H. 
Published: August 2005

AbstractIn this paper we develop four measures to describe the distribution of nuclear chromatin. These measures attempt to describe in an objective and meaningful way the heterogeneity, granularity, condensation, and margination of chromatin in cell nuclei. Starting with a high‐resolution digitized image of a cell where the nuclear pixels have been identified, the four measures may be rapidly estimated. The range of each is derived and the interpretation of the measures in the context of chromatin compaction and distribution is developed. Implementation issues such as sampling density, thresholding and subsequent pre‐processing, and algorithmic complexity are discussed.
Flow cytometry reveals a high degree of genomic size variation and mixoploidy in various strains of the acellular slime mold Physarum polycephalum
Authors: Kubbies M. , Wick R. , Hildebrandt A. , Sauer H. W. 
Published: August 2005

AbstractHigh‐resolution flow cytometry, using avian erythrocytes as an internal standard, was employed to study constitutive genome size variation of G2‐phase nuclei of Physarum polycephalum strains during the macroplasmodial stage of their life cycle. Our results document a previously unknown extent of genome size variation and mixoploidy in this organism. The unimodal diploid strain Tu 291 displayed the largest genome of the strains tested; in contrast, the Colonia strain displayed only half of the Tu 291 G2‐phase fluorescence, confirming its haploid nature. An additional strain, derived from a recent cross between Lu897 and Lu898 amoebae, must have arisen by selfing (propagation of only one of the parental genomes to the macroplasmodial stage), since its nuclei display close to the haploid G2‐phase DNA content. The observation of a small fraction of corresponding diploid nuclei within the haploid population of this strain, while maintained as microplasmodia, supports the notion that meiosis in haploid strains may require the presence of diploid nuclei. Two of the descendants of the prototype haploid Colonia strain, which were kept for extended periods of time in submerse culture, proved to be near diploid and mixoploid. Polyploidization and subsequent loss of DNA thus seems to contribute to the extremes of genome size variation in Physarum. In addition to unimodal fluorescence distributions, a number of diploid strains displayed bi‐ and even trimodal distributions within harvests of a single G2‐phase macroplasmodium. Analysis of these mixoploid strains by means of gaussian curve‐fitting suggests that the smaller genome size differences in Physarum may arise in step‐wise diminution of DNA in approximate units of 3–5% of the original Tu 291 genome.
A comment on “A method to measure the duration of DNA synthesis and the potential doubling time from a single sample”
Authors: Allen White R. , Meistrich Marvin L. 
Published: August 2005

AbstractRecently a method was introduced by Begg et al. (Cytometry 6:620–626, 1985) for the calculation of the DNA synthesis time, TS using flow cytometry to monitor the progression of cells that have been pulse‐labeled by bromodeoxyuridine (BrdUrd). Ideally, the method uses only a single two‐parameter histogram of the progressed cells to obtain TS. The essence of the method is to be able to obtain an initial point from general principles, draw a line through it and the measured point, and extrapolate to a level of progression that would be attained when the time is equal to TS. In this comment, explicit values are derived for the initial point and the progression of cells based on kinetic arguments, and a new method is presented for estimating TS. This new method is compared to earlier methods, and the effects of a finite labeling time and variation in the transit times are considered. Finally, an equation is developed for estimating TS following multiple measurements.
Thiazole orange: A new dye for reticulocyte analysis
Authors: Lee Linda G. , Chen Chia‐Huei , Chiu Laura A. 
Published: August 2005

AbstractThe purpose of this study was to find a 488‐nm excitable fluorescent dye for reticulocyte analysis by the use of fluorescence activated cell cytometry. The chemical structure of thioflavin T, a dye used for reticulocyte analysis with 457‐nm excitation, was used as a model. Several dyes were synthesized and evaluated by quantum yield determination, fluorescence microscopy, and flow cytometry. The best results were obtained with a dye we have named “thiazole orange”; analysis of several blood samples with thiazole orange gave a correlation coefficient of 0.97 as compared to a manual determination of reticulocyte percentage.
Segmentation of stained blood cell images measured at high scanning density with high magnification and high numerical aperture optics
Authors: Harms H. , Aus H. M. , Haucke M. , Gunzer U. 
Published: August 2005

AbstractIn hematological morphology, it is necessary to resolve and analyze the smallest possible cellular details appearing in the light microscope. A prerequisite for computer‐aided analysis of subtle morphological features is measuring the cells at a high scanning density with high magnification and high numerical aperture optics. Contrary to visual observations, the information content in a measured picture can be increased by setting the condensor's numerical aperture (NA) greater than the objective's NA. The complexity and heterogeneity of such cell images necessitate a new segmentation method that conserves the morphological information required in the subsequent image analysis, feature extraction, and cell classification. In our segmentation strategy, characteristic color difference thresholds for each nucleus and cytoplasm are combined with geometric operations, probability functions, and a cell model. All thresholds are repeatedly recalculated during the successive improvements of the image masks. None of the thresholds are fixed. This strategy segments blood cell images containing touching cells and large variations in staining, texture, size, and shape. Biological inconsistencies in the calculated cell masks are eliminated by comparing each mask with the cell model criteria integrated into the entire segmentation process. All 20,000 leukocyte images from 120 smears in our leukemia project were segmented with this method.
Random signal variation as a cause of systematic microdensitometric error
Authors: Goldstein D. J. 
Published: August 2005

AbstractA high noise/signal ratio in microdensitometry, owing, for example, to the use of a very small measuring spot or light at an extreme of the spectrum, decreases the precision (increases the variability) of results. This can be compensated for by making more measurements, but under some circumstances random signal variations can also cause systematic errors that are not so easily corrected. The present article explains how such errors can arise when measuring either very pale or very dense objects and suggests methods to detect and avoid them. The principles described are applicable to many types of microdensitometer, not only the Vickers flyingspot instruments used in this investigation.
Centrifugal separation of cells in sputum specimens from patients with undifferentiated carcinoma
Authors: Albright Craig D. , Frost John K. , Pressman Norman J. 
Published: August 2005

AbstractEthanol‐fixed cells in sputum from patients with undifferentiated carcinoma of the lung were separated in aqueous Ficoll using a discontinuous density gradient centrifugation technique. The selective enrichment of small cell undifferentiated (e.g., oat cell) or large cell undifferentiated carcinoma cells was achieved while removing most of the leukocytes (80–90%) and macrophages (65–75%) from specimen fractions containing the greatest relative frequencies of cancer cells. The maximum purity of small cell carcinoma cells (0.04%) occurs in moderate density (ρ = 1.121g/ml) gradient fractions and results in a 2.4‐fold enrichment relative to unprocessed specimens. In contrast, the maximum purity of large cell carcinoma cells (0.22%) is obtained in very high density (ρ = 1.172 g/ml) gradient fractions and results in a 1.2‐fold enrichment in comparison with unprocessed specimens. Microscopic examination of Papanicolaoustained specimen fractions reveals that these enrichments were achieved while retaining diagnostically significant cytomorphologic and tinctorial features necessary for cancer screening and diagnosis. Peak purity ranges of undifferentiated cancer cells significantly overlap comparable ranges for material from bronchogenic adenocarcinoma and squamous cell carcinoma.
The fate of the primary diploid population during spontaneous transformation of growth factor‐supplemented murine cell cultures
Authors: Kubbies M. , Friedl R. , Bettecken T. , Hoehn H. 
Published: August 2005

AbstractPrimary cultures derived from lung and renal tissue of the newborn harvest mouse (Micromys minutus) were serially passaged in media supplemented with epidermal growth factor, hydrocortisone, transferrin, insulin, and triiodothyronine. Although these growth factor supplements eliminated the growth crisis commonly encountered during the initial stages of murine primary cultures, the original diploid cell fraction clearly underwent such a “crisis”; the truly diploid cells invariably disappeared as these cultures reached 20 to 40 population doublings. They were replaced, either gradually or precipitously, by various heteroploid cell fractions. In three of four independent cultures, these “established” cells were hypotetraploid and appeared to be derived from a small number of progenitors already present during the very early (precrisis) culture stages. In contrast to rather frequent DNA changes displayed by clones and subclones derived from the various heteroploid cell lineages, the predominant components of the established mass cultures displayed a highly constant DNA fluorescence pattern. Our results suggest that primary murine cell cultures develop heteroploid cell lineages even if the initial growth crisis is mitigated by growth factor supplements. These heteroploid cells appear to respond more efficiently to stimulation by various growth factors than the primary diploid cell population.
Cell‐by‐cell autofluorescence correction for low signal‐to‐noise systems: Application to epidermal growth factor endocytosis by 3T3 fibroblasts
Authors: Roederer Mario , Murphy Robert F. 
Published: August 2005

AbstractAutofluorescence of cells can be a major portion of the fluorescence signal in many systems, especially when fluorescent conjugates are used to study receptor‐ligand systems for which there are less than 70,000 receptors per cell. We have devised a method for the cell‐by‐cell correction of autofluorescence for flow cytometric data by using an additional parameter to measure and correct for autofluorescence in the fluorescence channel. The principle has been extended to allow simultaneous correction for autofluorescence and dual‐fluorescence spillover compensation in samples labeled with two different fluorochromes; all corrections were done in software, making them applicable to any flow cytometer. The autofluorescence correction method was used to analyze the acidification of epidermal growth factor (EGF) by Swiss 3T3 cells. EGF is acidified to pH 6.2 starting two min after labeling, with a half‐time for acidification of 45 s.
Orientation measurements of microsphere doublets and metaphase chromosomes in flow
Authors: Lucas Joe Nathan , Pinkel Daniel 
Published: August 2005

AbstractObtaining information about the shape of particles from slit‐scan profiles is facilitated if the particles are oriented. Elongated particles orient in the nozzle of flow cytometers, but orientation may be disrupted before the particles get to the point of measurement. We have used our slit‐scan flow cytometer to investigate the orientation of microsphere doublets in a liquid jet in air, in flow across a glass surface, and in a 200‐μm‐square capillary tube as a function of distance from the flow chamber nozzle. Particles were classified as being oriented if there was a centrally located dip in the slit‐scan profile. Essentially all the doublets in the jet were oriented, and no disorientation was noted over the distances measured (up to 10 mm from the nozzle). Particle orientation was maintained for 80 μm in flow across a glass surface. In the capillary‐type flow chamber, essentially all of the particles were oriented at the tube entrance and for several millimeters into the tube. There then occurred a region where particle tumbling started and progressively fewer doublets met the orientation criteria. The distance to where tumbling began could be estimated by calculating the length required to establish the parabolic flow profile in the tube. Finally, the fraction of oriented particles reached a constant value that did not change with increased distance into the tube. When sample was injected off axis (i.e., half‐way between the chamber center and the chamber walls), particle tumbling began closer to the tube entrance.Orientation studies of Chinese hamster M3–1 metaphase chromosomes showed that over 95% of the larger chromosomes (1–5) and over 65% of the smaller chromosomes (9–11) were oriented near the entrance of the capillary tube.
Swine chromosomes: Flow sorting and spot blot hybridization
Authors: Grunwald D. , Geffrotin C. , Chardon P. , Frelat G. , Vaiman M. 
Published: August 2005

AbstractFlow cytometry analysis was applied to swine chromosomes prepared from phytohemagglutinin (PHA) stimulated peripheral blood lymphocytes. Flow karyotypes from both sexes and from t(3;7) translocation carrier females were obtained. A certain number of chromosome pairs could be assigned to various peaks. In fact, 13 peaks were observed for 18 autosomal pairs plus X and Y. Moreover, abnormalties owing to the t(3;7) translocation were readily observable.The number of base pairs for chromosomes associated with the various peaks was estimated by comparison with human flow karyotypes.The following four peaks were thus sorted: (1) the peak assumed to represent the translocated chromosome 7 plus the normals associated with it; (2) the corresponding peak from a normal swine; (3) the peak assumed to contain among others the normal chromosome 7; and finally (4) the peak corresponding to swine chromosome 1.Chromosomes of each peak were collected on Pall Biodyne membrane. Following appropriate denaturation and prehybridization, the four samples were hybridized with a human leucocyte antigen (HLA) class I32P‐labelled cDNA probe, representing most of the coding sequence of the HLA B7 gene. The results confirmed previous data from other techniques that assigned the swine MHC(SLA) to chromosome 7.Subsequently, sorted samples were hybridized with a porcine genomic Interferon alpha probe in order to confirm the mapping of this gene family on porcine chromosome 1.