Optical errors in scanning stage absorbance cytophotometry. I. Procedures for correcting apparent integrated absorbance values for distributional, glare, and diffraction errors.

Duijndam, W. A. L.; Smeulders, A.W.M.; Duijn, Van; Verweij, A.C.

doi:10.1177/28.5.6991589

Cited by 43 publications

(5 citation statements)

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“…(4) Adjust "offset" and "gain" before each measurement. [4,16,20,21,28,29,36,37,41,44,48,50]. Corresponding quality assurance protocols are given in Part II (Quality control of ICM instrumentation).…”

Section: B2 Instrumentation Requirements For Densitometric Measuremmentioning

confidence: 99%

Part I: Basic Considerations and Recommendations for Preparation, Measurement and Interpretation

Haroske

Giroud

Reith

et al. 1998

Analytical Cellular Pathology

111

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A task force of invited experts in the field of diagnostic DNA image cytometry, especially consisting of participants from the PRESS (Prototype Reference Standard Slides) and EUROPATH (European Pathology Assisted by Telematics for Healthcare) projects, but open to any other scientist or physician revealing experience in that new diagnostic procedure (names are given in the Annex A) agreed upon the following updated consensus report during the 5th International Congress of the ESACP 1997 in Oslo. This report is based on the preceeding one [9] and on results of the above mentioned European research projects. It deals with the following items: – Biological background and aims of DNA image cytometry,– Principles of the method,– Basic performance standards,– Diagnostic interpretation of DNA measurements,– Recommendations for practical use.

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Section: B2 Instrumentation Requirements For Densitometric Measuremmentioning

confidence: 99%

Part I: Basic Considerations and Recommendations for Preparation, Measurement and Interpretation

Haroske

Giroud

Reith

et al. 1998

Analytical Cellular Pathology

111

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“…Although this possibility cannot be completely ruled out, we think it is unlikely because it is not supported by any available evidence. In particular, no known factor in the DNA structure, conformation, or compactness can account for a 2-fold difference in the Feulgen-reaction fluorescence intensity (8,(15)(16)(17). In addition, the microscopic examination of the chromatin in metacyclic trypanosomes shows no trace of pycnosis or of any other feature that could account for this difference in fluorescence.…”

Section: Resultsmentioning

confidence: 91%

Evidence for haploidy in metacyclic forms of Trypanosoma brucei.

Zampetti‐Bosseler¹,

Schweizer²,

Pays³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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The parasitic flagellate Trypanosoma brucei undergoes a series of morphologic and metabolic changes during its passage in the digestive organs of its insect vector, a Glossina or tsetse fly. This morphogenesis ends by the differentiation, in the salivary gland of the fly, of the metacyclic form, which will be transmitted in the bloodstream of the mammalian host. On the basis of DNA microfluorometric measurements, we propose that these metacyclic trypanosomes have a haploid amount of DNA, compared to that of bloodstream forms and also of the proventricular forms, which initiate the invasion of the salivary glands. It can be inferred that trypanosomes undergo meiosis during their developmental cycle in the tsetse fly's salivary glands and syngamy shortly after cyclic transmission.During its life cycle, Trypanosoma brucei differentiates into a series of cytologically and metabolically distinct forms. The principal forms identified in the blood of the mammalian host are the slender form, which rapidly divides and can undergo antigenic variation, and the stumpy, nondividing form. After the tsetse fly has fed on blood from an infected mammalian host, the ingested trypanosomes differentiate into the procyclic and proventricular forms in the digestive tract and subsequently into the epimastigote and metacyclic forms in the salivary glands of the fly. Although many aspects of this parasitic cycle are now well documented (see ref. 1 for a recent review), the existence of a sexual cycle in trypanosomes has been and still remains an open question (2). The identification of mitosis, meiosis, and the ploidy of these parasites is not possible by direct cytogenetic analysis because their genome does not condense into defined chromosomes at any stage of the cell cycle. However, evidence based on isoenzyme studies (2, 3) and on the recent demonstration of hybrid formation between two clones of T. brucei (4) shows that genetic exchange does take place in trypanosomes. Analysis of restriction site polymorphisms (5) and direct measurements of DNA content (6) and genome complexity (7) indicate that T. brucei is diploid. Borst et al. (6) measured the nuclear DNA content of T. brucei using quantitative absorption and fluorescence cytophotometry of individual Feulgen reaction-pararosaniline-stained cells. They performed their analyses on bloodstream and procyclic culture forms and concluded that both forms have the same DNA content. However, the amount of DNA has never been measured in trypanosomes during their developmental cycle in the tsetse fly vector. Therefore, we estimated by direct microfluorometry the nuclear DNA content, after Feulgen reaction-pararosaniline staining of proventricular and metacyclic trypanosomes. These forms are found in the saliva of flies just before and after development of the parasite in the salivary glands, respectively. The quantitative cytochemical method we used is based on the observation of a linear relationship between the amount of fluorescence and the DNA content of Feulgen-stained nucl...

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“…It is this variance that mainly defines the degree of accuracy that can be reached in a DNA ploidy analysis, as it was already demonstrated in a previous paper [17]. All methodological and technological improvements [9,11,12,16,19,26] of the measurement process which lead to an increased precision will also reduce this very important error source, and will therefore improve the accuracy.…”

Section: Discussionmentioning

confidence: 93%

DNA Histogram Interpretation Based on Statistical Approaches

Haroske

Dimmer²,

Meyer³

et al. 1997

Analytical Cellular Pathology

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Image cytometric DNA measurements provide data which are most often interpreted as equivalent to the chromosomal ploidy although the chromosomal and the DNA ploidy are not identical. The common link between them is the cell cycle. Therefore, if destined for DNA ploidy interpretations, the DNA cytometry should be performed on a population‐oriented stochastic basis. Using stochastic sampling the data can be interpreted by applying the rules of stochastic processes. A set of statistical methods is given that enables a DNA histogram to be interpreted objectively and without human interaction. These statistics analyse the precision and accuracy of the entire measurement process. They give in error probabilities for accepting a measurement as reliable, for recognition of stemlines, stemline aneuploidy, and for evaluating so‐called rare events. Nearly 300 image cytometric DNA measurements from breast cancers and rat liver imprints examples have been selected to demonstrate the efficiency of the statistics in each step of interpreting DNA histograms.

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Optical errors in scanning stage absorbance cytophotometry. I. Procedures for correcting apparent integrated absorbance values for distributional, glare, and diffraction errors.

Cited by 43 publications

References 0 publications

Part I: Basic Considerations and Recommendations for Preparation, Measurement and Interpretation

Part I: Basic Considerations and Recommendations for Preparation, Measurement and Interpretation

Evidence for haploidy in metacyclic forms of Trypanosoma brucei.

DNA Histogram Interpretation Based on Statistical Approaches

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