&lt;title&gt;Automated microscopy system for detection and genetic characterization of fetal nucleated red blood cells on slides&lt;/title&gt;

Ravkin, Ilya; Temov, Vladimir

doi:10.1117/12.307091

Cited by 15 publications

(5 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The scanner used for detection of antibody-stained NRBCs and FISH signals has been developed by Applied Imaging Corp. [11,12]. The instrument is a microscope-based scanner equipped with a oneslide motorized stage and a black-and-white video camera.…”

Section: Identification and Acquisition Of Fish Signals By Scanningmentioning

confidence: 99%

Sensitivity and Specificity of the Identification of Fetal Cells in Maternal Blood by Combined Staining with Antibodies against Beta-, Gamma- and Epsilon-Globin Chains

et al. 2003

View full text Add to dashboard Cite

Objectives: Antibodies against fetal and embryonic hemoglobins may identify fetal cells in maternal blood. Both γ- and Ε-globins are used as fetal cell markers. γ-Globin is not fetus specific. So far Ε-globin has been claimed to be fetus specific. In this communication, we compare the specificity of anti-Ε- and anti-γ-globin staining when combined with staining for β-globin. Methods: We applied single and double color immunofluorescent staining techniques in combination with XY chromosome hybridization. The blood sample was taken after chorion villus biopsy at 11 weeks of gestation from a woman carrying a male fetus. Results: By γ-globin staining alone, 21 fetal and 2 maternal nucleated red blood cells (NRBCs) were identified. Only 1 of the 2 maternally derived NRBCs expressed β-globin. By Ε-globin staining, 92 additional fetal NRBCs were identified. Conclusions: Ε-Globin antibody and combined Ε- and γ-globin antibody staining of a blood sample from a pregnant woman at 11 gestational weeks showed higher sensitivity but lower specificity for the fetal origin of erythroblasts with combined compared with separate staining. The final decision of the origin of cells was made by gender determination by FISH. Out of 2 γ-positive maternal cells 1 was β-globin antibody positive, 1 was β-globin negative, indicating that 100% specificity for fetal origin could not be obtained by combining all 3 hemoglobin types. Although only 1 blood sample was tested and only 2 γ-positive maternal NRBCs were identified, the result indicates that β-hemoglobin does not discriminate completely between γ-positive NRBCs of fetal and maternal origin.

show abstract

Section: Identification and Acquisition Of Fish Signals By Scanningmentioning

confidence: 99%

Sensitivity and Specificity of the Identification of Fetal Cells in Maternal Blood by Combined Staining with Antibodies against Beta-, Gamma- and Epsilon-Globin Chains

et al. 2003

View full text Add to dashboard Cite

show abstract

“…The consequence is a wide variation in results reported by different observers of the same blood smears. 5 To address this issue, Cupp and colleagues, 45 Medearis and colleagues, 46 Ravkin and Temov, 47 and Pelikan and colleagues 48,49 proposed automated laboratory systems designed to increase the number of RBCs scanned and improve the objectivity of end point determinations. The Pelikan automated computer-assisted microscopy system consisted of an automated scanning stage, microsetting motor controls, and bright-field analysis using green and red absorption filters.…”

Section: Modified K-b Acid-elution Assay: Automated Detectionmentioning

confidence: 99%

Laboratory methods for Rh immunoprophylaxis: a review

Sandler¹,

Sathiyamoorthy

2010

Immunohematology

View full text Add to dashboard Cite

The recommended dose of Rh immune globulin for postpartum Rh immunoprophylaxis is based on an estimation of the volume of the fetomaternal hemorrhage, if any, measured as the percent of fetal RBCs in a sample of the D-mother's blood. Laboratory methods for distinguishing fetal from maternal RBCs have been based on their different blood types (D+ versus D-) or predominant hemoglobin content (hemoglobin F versus hemoglobin A). We conducted a review of the medical literature describing laboratory methods for detecting and quantifying fetal RBCs in maternal blood samples. We also used data collected for the College of American Pathologists Fetal RBC Detection Surveys to determine which laboratory methods are used currently in hospitals in the United States. The rosette screen is used widely for identifying D-mothers who may require additional doses of Rh immune globulin for postpartum immunoprophylaxis. As the rosette screen targets the D antigen, it is not suitable for detecting a fetomaternal hemorrhage in D+ mothers or when the D type of the fetus or newborn is D-or unknown. The acid-elution (Kleihauer-Betke) assay is a sensitive laboratory method for quantifying a fetomaternal hemorrhage, but it is tedious, often inaccurate, and difficult to reproduce. Flow cytometry, using anti-D or antihemoglobin F reagents, offers a more precise quantification of fetal RBCs in maternal blood. However, flow cytometry services for this function are available in relatively few hospital laboratories in the United States because of logistic and fiscal impediments.

show abstract

“…This separation yields 20–30 million mononuclear cells. Because this is still a large number, additional enrichment steps such as cell lysis (8) and magnetic activated cell sorting (MACS) (9, 10) are commonly employed. These methods are often used in series, resulting in a time‐ and labor‐intensive process with unavoidable losses in the target cell population at each step.…”

Section: Reported Flow and Image Cytometry System Performancementioning

confidence: 99%

Ultra-rare-event detection performance of a custom scanning cytometer on a model preparation of fetal nRBCs

et al. 2000

View full text Add to dashboard Cite

Background The performance of a fully automated scanning cytometer incorporating previously reported high‐precision autofocus and accurate image segmentation was evaluated for the detection of “ultra‐rare” cells using a model of fetal nucleated red blood cells (fnRBCs) in the maternal circulation. These distinctive scanning cytometry techniques were expected to markedly improve sensitivity and specificity. Methods Normal adult blood and fetal red blood cells were stained with fluorescein isothiocyanate–conjugated anti‐fetal hemoglobin and 4,6‐diamidino‐2‐phenylindole, a nuclear dye. Adult cells were spiked with fetal cells to create ratios of about 1 fnRBC in 107 nucleated cells and deposited in monolayers on slides using centrifugal cytology. Rare‐event performance parameters were reviewed, formalized, and applied to test the new instrument using this cell model. Results Fifteen slides were analyzed to establish performance by comparison with manual detection, and four sets of four slides each were then scanned to explore the limit of detection. Results were an average sensitivity of 91%, an average specificity error of 12.3 false‐positives per million cells, and repeatability of 100% at a cell analysis rate of 862 Hz. With addition of a quick interactive step subsequent to scanning, the false‐positive rate dropped to a total of only one artifact over the 15 experiments. The instrument succeeded at locating 1 fnRBC in 20 million adult cells, the lowest limit of detection tested. Conclusion This consistently high performance, coupled with the capability of scanning arbitrarily large numbers of cells, validates the considerable potential of precise high‐speed autofocus and accurate real‐time image segmentation for ultra‐rare event detection. Cytometry 39:285–294, 2000 © 2000 Wiley‐Liss, Inc.

show abstract

<title>Automated microscopy system for detection and genetic characterization of fetal nucleated red blood cells on slides</title>

Cited by 15 publications

References 12 publications

Sensitivity and Specificity of the Identification of Fetal Cells in Maternal Blood by Combined Staining with Antibodies against Beta-, Gamma- and Epsilon-Globin Chains

Sensitivity and Specificity of the Identification of Fetal Cells in Maternal Blood by Combined Staining with Antibodies against Beta-, Gamma- and Epsilon-Globin Chains

Laboratory methods for Rh immunoprophylaxis: a review

Ultra-rare-event detection performance of a custom scanning cytometer on a model preparation of fetal nRBCs

Contact Info

Product

Resources

About