FISHprep: A Novel Integrated Device for Metaphase FISH Sample Preparation

Shah, Pranjul Jaykumar; Vedarethinam, Indumathi; Kwasny, Dorota; Andresen, Lars Ole; Skov, Søren; Silahtaroglu, Asli; Tümer, Zeynep; Svendsen, Winnie Edith

doi:10.3390/mi2020116

Cited by 13 publications

(10 citation statements)

References 43 publications

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“…The quality of spreads obtained from the FISHprep was comparable to traditionally obtained spreads. The integration of all the steps required for successful metaphase FISH slide preparation in one FISHprep offers a possibility for an automation of the molecular cytogenetics in future (Shah et al, 2011b). …”

Section: Integrated Culturing Harvesting and Splashing Devicementioning

confidence: 99%

Microtechnologies Enable Cytogenetics

Kwasny¹,

Vedarethinam²,

Shah³

et al. 2012

Recent Trends in Cytogenetic Studies - Methodologies and Applications

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Section: Integrated Culturing Harvesting and Splashing Devicementioning

confidence: 99%

Microtechnologies Enable Cytogenetics

Kwasny¹,

Vedarethinam²,

Shah³

et al. 2012

Recent Trends in Cytogenetic Studies - Methodologies and Applications

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“…This procedure involved treatment of cells with a hypotonic salt solution, fixation of the suspended cell pellet, and dropping of the cells onto glass slides. Svendsen et al [16][17][18] introduced a novel splashing device for preparing metaphase spreads on a microscope glass slide. They reported the application of a semiclosed microfluidic chip to fast evaporation of the fixative solution.…”

Section: ·3 Operationmentioning

confidence: 99%

“…[16][17][18] To obtain a clearly visible image, metaphase chromosomes were spread and fixed on the slide in a manner to reduce the overlap of chromosomes. This procedure involved treatment of cells with a hypotonic salt solution, fixation of the suspended cell pellet, and dropping of the cells onto glass slides.…”

Section: ·3 Operationmentioning

confidence: 99%

Microdevice in Cellular Pathology: Microfluidic Platforms for Fluorescence in situ Hybridization and Analysis of Circulating Tumor Cells

Sato

2015

ANAL. SCI.

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867 IntroductionCellular pathology techniques have progressed in recent years. Conventionally, the pathologist observes tissues with a microscope and determines whether or not the hematoxylin and eosin (H&E)-stained tissues contain cancer cells based on their morphology. The H&E stain is the traditional and most widely used staining method in medical diagnoses. 1 Recent advances in genetic analysis methods have made it possible to identify the genetic basis of human diseases, and have opened the door to individualized prevention strategies and early detection and treatment.Therefore, combinations of the traditional morphological diagnosis method and newer genetic methodologies are strongly welcomed.Fluorescence in situ hybridization (FISH) is a well-known gene-based method used to image genetic abnormalities. FISH is a microscopic technique in which specific DNA sequences tagged with fluorophores are used to detect target genes and identify their localization within a cell. FISH was developed in the early 1980s 2 and has since been improved continuously. 3 The FISH probes can only detect genes with a high degree of homology, and can visualize specific cytogenetic abnormalities such as copy number aberrations, gene mutations, and gene fusions. 4 This advanced molecular pathology technique has enabled better diagnoses of diseases, leading to more tailored therapeutic regimens.Analytes have become more multifaceted. Microfluidic devices enable the miniaturization, integration, automation, and parallelization of chemical and biochemical processes. This new technology also provides opportunity for expansion in the field of cellular pathology. Fluorescence in situ hybridization (FISH) is a well-known gene-based method to image genetic abnormalities. Development of a FISH microfluidic platform has offered the possibility of automation with significant time and cost reductions, which overcomes many drawbacks of the current protocols. Microfluidic devices are also powerful tools for single-cell analysis. Capturing the circulating tumor cells (CTCs) from blood samples is one of the most promising approaches to enable the early diagnosis of cancer. The microfluidic devices are also useful to isolate rare CTCs at high efficiency and purity. In this review, I outline recent FISH and CTC analyses using microfluidic devices, and describe their applications for the cellular diagnosis of cancers.

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“…The FISH is applied on metaphase chromosomes, interphase nuclei, ixed tissues or cells and solid tumour samples [22]. The procedure contains denaturation, hybridization, post hybridization washes, which removes unbound single-strand DNA and after washing, an anti-fade solution containing DAPI (4′,6-diamidino-2-phenylindole) is applied to the slide, and a coverslip must be added ( Figure 1) [23]. For FISH analysis, epiluorescence microscopes with speciic ilters and for identifying luorochromes, a charge-coupled device (CCD) camera that captures the images were needed.…”

Section: Fish Developmentmentioning

confidence: 99%

The Use of Molecular Cytogenetic Techniques for the Identification of Chromosomal Abnormalities

Kalkan¹

2017

Chromosomal Abnormalities - A Hallmark Manifestation of Genomic Instability

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Chromosomal analysis is an increasingly important diagnostic procedure in numerous areas of clinical medicine that includes haematology, perinatology or obstetrics. Chromosomal disorders are viewed as a major category of genetic diseases, and sometimes the identiication of abnormal chromosomes is not easily applicable. Just like the identiication of the marker chromosome or the identiication of the complex karyotypes is important in clinics for the evaluation of the patient prognosis as well as the treatment response, needless to say; luorescence in situ hybridization (FISH) is the most suitable and rapid method in the above-mentioned situations. It gives chance to the rapid analysis of chromosomal aneuploidies in dividing and non-dividing cells. In this chapter, we will discuss the general principles of the chromosomal abnormalities and the molecular cytogenetic techniques that can help the identiication of presence or absence of a particular DNA sequence or the evaluation of the number of organization of a chromosome or chromosomal region.

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FISHprep: A Novel Integrated Device for Metaphase FISH Sample Preparation

Cited by 13 publications

References 43 publications

Microtechnologies Enable Cytogenetics

Microtechnologies Enable Cytogenetics

Microdevice in Cellular Pathology: Microfluidic Platforms for Fluorescence in situ Hybridization and Analysis of Circulating Tumor Cells

The Use of Molecular Cytogenetic Techniques for the Identification of Chromosomal Abnormalities

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