A Semi-Closed Device for Chromosome Spreading for Cytogenetic Analysis

Kwasny, Dorota; Mednova, Olga; Vedarethinam, Indumathi; Silahtaroglu, Asli; Tümer, Zeynep; Almdal, Kristoffer; Svendsen, Winnie Edith

doi:10.3390/mi5020158

Cited by 6 publications

(9 citation statements)

References 24 publications

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“…To reduce the workflow for FISH, microfluidics has been suggested by several groups with the processing time reduced from several days using conventional slide-based FISH to several hours using FISH-on-chip platforms [14,[21][22][23]. In addition, microfluidic platforms have also resulted in a reduction of the use of expensive FISH probes.…”

Section: Discussionmentioning

confidence: 99%

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Microfluidic Device for On-Chip Immunophenotyping and Cytogenetic Analysis of Rare Biological Cells

Weerakoon-Ratnayake¹,

Vaidyanathan²,

Larkey³

et al. 2019

Preprint

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The role of circulating plasma cells (CPCs) and circulating leukemic cells (CLCs) as biomarkers for several blood cancers, such as multiple myeloma and leukemia, respectively, have recently been reported. These markers can be attractive due to the minimally invasive nature of their acquisition through a blood draw (i.e., liquid biopsy) negating the need for painful bone marrow biopsies. CPCs or CLCs can be used for cellular/molecular analyses, such as immunophenotyping or fluorescence in situ hybridization (FISH). FISH, which is typically carried out on slides involving complex workflows, becomes problematic when operating on CLCs or CPCs due to their relatively modest numbers. Here, we present a microfluidic device for characterizing CPCs and CLCs enriched from peripheral blood using immunofluorescence or FISH. The microfluidic possessed an array of cross-channels (2-4 µm in depth and width) that interconnected a series of input and output fluidic channels. Placing a cover plate over the device formed microtraps, the size of which was defined by the width and depth of the cross-channels. This microfluidic chip allowed for automating immunofluorescence and FISH requiring the use of small volumes of reagents, such as antibodies and probes, as compared to slide-based immunophenotyping and FISH. In addition, the device could secure FISH results in <4 h compared to 2-3 d for conventional FISH.

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Section: Discussionmentioning

confidence: 99%

“…Kwasny et al [21] described the use of cyclic olefin copolymer (COC) devices sealed with glass or COC cover plates that were surface-modified to allow for stretching chromosomes. Perez-Torella and coworkers reported a COC chamber, which was capable of delivering FISH reagents to cells [22].…”

Section: Introductionmentioning

confidence: 99%

Microfluidic Device for On-Chip Immunophenotyping and Cytogenetic Analysis of Rare Biological Cells

Weerakoon-Ratnayake¹,

Vaidyanathan²,

Larkey³

et al. 2019

Preprint

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“…Shah and coworkers built a device, which integrates multiple stages of the process, allowing for initial culturing, arrest, and fixation of metaphase cells as well as having the ability to prepare metaphase chromosome spreads on glass slides for metaphase FISH analysis (Kwasny, et al, 2012). Creating the chromosome spreads has been described as more of an art than a science, but many devices (including the device built by Shah and co-workers) are being built which make the creation of these spreads more reliable and repeatable (Kwasny, et al, 2012;Kwasny, et al, 2014). But al., 2014) the system relied on a conventional thermal cycler, power supply (for DEP) and fluorescence microscope, potentially impacting its portability for POC use.…”

Section: Microfluidic Sprep In Cytogeneticsmentioning

confidence: 99%

Advances in Microfluidics and Lab-on-a-Chip Technologies

Jayamohan

Romanov

et al. 2017

Molecular Diagnostics

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Advances in molecular biology are enabling rapid and efficient analyses for effective intervention in domains such as biology research, infectious disease management, food safety, and biodefense. The emergence of microfluidics and nanotechnologies has enabled both new capabilities and instrument sizes practical for point-of-care. It has also introduced new functionality, enhanced sensitivity, and reduced the time and cost involved in conventional molecular diagnostic techniques. This chapter reviews the application of microfluidics for molecular diagnostics methods such as nucleic acid amplification, next-generation sequencing, high resolution melting analysis, cytogenetics, protein detection and analysis, and cell sorting. We also review microfluidic sample preparation platforms applied to molecular diagnostics and targeted to sample-in, answer-out capabilities

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“…[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%

“…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%

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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A Semi-Closed Device for Chromosome Spreading for Cytogenetic Analysis

Cited by 6 publications

References 24 publications

Microfluidic Device for On-Chip Immunophenotyping and Cytogenetic Analysis of Rare Biological Cells

Microfluidic Device for On-Chip Immunophenotyping and Cytogenetic Analysis of Rare Biological Cells

Advances in Microfluidics and Lab-on-a-Chip Technologies

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

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