Microscopy and Image Analysis

McNamara, George; Difilippantonio, Michael J.; Ried, Thomas

doi:10.1002/0471142905.hg0404s46

Cited by 15 publications

(12 citation statements)

References 151 publications

(156 reference statements)

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“…A TORCH screen was carried out (by ARCHITECT i 1000SR, Abbott Diagnostics, USA) in cases of suspected congenital infection. Chromosomal assays [ 13 , 14 ] were performed in cases of suspected chromosomal abnormities. Neonates suspected of having NAT, based on the presentation and clinic course of the illness, were designated as having an idiopathic cause as the diagnostic test for NAT is not available in our laboratory.…”

Section: Methodsmentioning

confidence: 99%

Risk factors for severity of thrombocytopenia in full term infants: a single center study

et al. 2021

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Background Neonatal thrombocytopenia (NT) (platelet count < 150 × 109/L) is a common finding in the neonatal intensive care unit (NICU). The main aim of this study was to assess the prevalence, risk factors, and outcomes of severe NT in full term (FT) infants. Methods During the study period, all FT infants who met the inclusion criteria for NT on two occasions were included. Maternal data, such as maternal age, weight, gestational age, mode of delivery, and history of systemic diseases, including diabetes mellitus, pre-eclampsia, systemic lupus erythematosus, and immune thrombocytopenic purpura, were recorded. Furthermore, neonatal data, such as gender, neonatal weight, causes/duration of admission, types of respiratory support used, complete blood count measurements, and outcomes for neonates admitted to the NICU, were recorded. Results In total, 55 FT infants with NT met the inclusion criteria, and 29 (52.73%) cases had severe NT. The most common cause of NT was neonatal sepsis (20 cases, 36.35%), followed by a postoperative state (5 cases, 9.09%). Moreover, in cases of positive blood cultures, the most commonly isolated organism was Escherichia coli (6 cases, 10.90%), followed by Klebsiella (5 cases, 9.09%). Cases of severe NT needed more platelet transfusions (P = 0.001) and had higher rates of mortality (P = 0.001) when compared to cases of mild/moderate NT associated with signs of bleeding and pulmonary/intraventricular hemorrhage (IVH) (P = 0.001). Conclusion Severe NT compared to mild/moderate NT, associated with signs of bleeding and pulmonary/IVH, needed more platelet transfusions, and had increased mortality. Further research is needed to explain which of these complications related to severity of thrombocytopenia or were associated with original disease of the babies.

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Section: Methodsmentioning

confidence: 99%

Risk factors for severity of thrombocytopenia in full term infants: a single center study

et al. 2021

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“…no. 59010, Chroma Technology Corp.; see also UNIT 4.4;McNamara et al, 2005) Aqua/Green/Red triple band (e.g., cat. no.…”

Section: Methodsmentioning

confidence: 99%

“…no. 69008, Chroma Technology Corp.; see also UNIT 4.4; McNamara et al, 2005) Microtome 65°C and 95°C ovens (or a slide warmer; e.g., HYBrite, Abbott Molecular, Leica Biosystems) 50-ml glass Coplin jars Epifluorescence microscope equipped with 100-W mercury lamp (UNIT 4.4; McNamara et al, 2005) 20× objective and 40× or 100× oil immersion fluorescence objectives 22 × 22-mm glass coverslips Humidified hybridization chamber (UNIT 4.3; Knoll and Lichter, 2005) 24 × 50-mm glass no.1 coverslips Additional reagents and equipment for paraffin embedding and preparation of tissue sections (Zeller, 1989), FISH (UNIT 4.3; Knoll and Lichter, 2005), and fluorescence microscopy (UNIT 4.4; McNamara et al, 2005) Prepare slides 1. Using a microtome, cut 4-to 6-μm sections of paraffin-embedded tissue and place on silanized glass microscope slides by floating the sections in water.…”

Section: Methodsmentioning

confidence: 99%

Preparation of Cells from Formalin‐Fixed, Paraffin‐Embedded Tissue for Use in Fluorescence In Situ Hybridization (FISH) Experiments

Weremowicz

2015

CP Human Genetics

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Numerical and structural chromosome abnormalities can be accurately detected in cells from archived tissues using fluorescence in situ hybridization (FISH). This unit describes two common approaches to performing FISH in formalin-fixed, paraffin-embedded tissue. The first approach utilizes 4 to 6 μm tissue sections in cases for which preserving tissue morphology is necessary, and the second involves extraction of intact nuclei from 50-μm tissue sections. To interpret FISH results using 4 to 6 μm sections, an adequate number of nuclei must be evaluated to perform statistical analysis. Evaluation of 30 to 50 nuclei from the single-cell suspension generally gives an interpretable result.

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“…3 and ref. 36). The epifluorescence microscope should be equipped with a ×63 oil-immersion objective, a single custom-designed triple band-pass filter (SKY filter cube) that allows for the simultaneous excitation of all fluorochromes, and a DAPI filter cube.…”

Section: ? Troubleshootingmentioning

confidence: 99%

Spectral karyotyping analysis of human and mouse chromosomes

et al. 2006

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Classical banding methods provide basic information about the identities and structures of chromosomes on the basis of their unique banding patterns. Spectral karyotyping (SKY), and the related multiplex fluorescence in situ hybridization (M-FISH), are chromosome-specific multicolor FISH techniques that augment cytogenetic evaluations of malignant disease by providing additional information and improved characterization of aberrant chromosomes that contain DNA sequences not identifiable using conventional banding methods. SKY is based on cohybridization of combinatorially labeled chromosome-painting probes with unique fluorochrome signatures onto human or mouse metaphase chromosome preparations. Image acquisition and analysis use a specialized imaging system, combining Sagnac interferometer and CCD camera images to reconstruct spectral information at each pixel. Here we present a protocol for SKY analysis using commercially available SkyPaint probes, including procedures for metaphase chromosome preparation, slide pretreatment and probe hybridization and detection. SKY analysis requires approximately 6 d.

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Microscopy and Image Analysis

Cited by 15 publications

References 151 publications

Risk factors for severity of thrombocytopenia in full term infants: a single center study

Risk factors for severity of thrombocytopenia in full term infants: a single center study

Preparation of Cells from Formalin‐Fixed, Paraffin‐Embedded Tissue for Use in Fluorescence In Situ Hybridization (FISH) Experiments

Spectral karyotyping analysis of human and mouse chromosomes

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