The Use of Microarrays and Fluorescence In Situ Hybridization for the Study of Mechanotransduction from Topography

McNamara, Laura E.; Dalby, Matthew J.; Tsimbouri, Monica P.

doi:10.1016/b978-0-12-416742-1.00015-9

Cited by 6 publications

(6 citation statements)

References 22 publications

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“…It is a well-known fact that the cells respond differently to the surfaces of different materials [86][87][88].…”

Section: Cellular Behavior As An Alternative Approach For the Previewmentioning

confidence: 99%

Cellular behavior as a dynamic field for exploring bone bioengineering: A closer look at cell–biomaterial interface

Gemini-Piperni

Takamori²,

Sartoretto

et al. 2014

Archives of Biochemistry and Biophysics

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“…It is a well-known fact that the cells respond differently to the surfaces of different materials [86][87][88].…”

Section: Cellular Behavior As An Alternative Approach For the Previewmentioning

confidence: 99%

Cellular behavior as a dynamic field for exploring bone bioengineering: A closer look at cell–biomaterial interface

Gemini-Piperni

Takamori²,

Sartoretto

et al. 2014

Archives of Biochemistry and Biophysics

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“…FISH offers numerous possibilities for studying either the whole genome or specific genomic loci (regions), and this technique has been widely used to detect aneuploidies and recurrent chromosomal abnormalities in preimplantation genetic, prenatal, and postnatal diagnoses and cancer cytogenetics. Moreover, the application of FISH has long been demonstrated as extremely valuable for studying chromosomal and genome organization, evolution and variations in health and disease (see Geurts and de Jong 2013; McNamara et al , 2014; Pita et al , 2014). …”

Section: Fish Applications In Pre- and Postnatal Diagnostics And Resementioning

confidence: 99%

Human molecular cytogenetics: from cells to nucleotides

Riegel

2014

Genet. Mol. Biol.

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The field of cytogenetics has focused on studying the number, structure, function and origin of chromosomal abnormalities and the evolution of chromosomes. The development of fluorescent molecules that either directly or via an intermediate molecule bind to DNA has led to the development of fluorescent in situ hybridization (FISH), a technology linking cytogenetics to molecular genetics. This technique has a wide range of applications that increased the dimension of chromosome analysis. The field of cytogenetics is particularly important for medical diagnostics and research as well as for gene ordering and mapping. Furthermore, the increased application of molecular biology techniques, such as array-based technologies, has led to improved resolution, extending the recognized range of microdeletion/microduplication syndromes and genomic disorders. In adopting these newly expanded methods, cytogeneticists have used a range of technologies to study the association between visible chromosome rearrangements and defects at the single nucleotide level. Overall, molecular cytogenetic techniques offer a remarkable number of potential applications, ranging from physical mapping to clinical and evolutionary studies, making a powerful and informative complement to other molecular and genomic approaches. This manuscript does not present a detailed history of the development of molecular cytogenetics; however, references to historical reviews and experiments have been provided whenever possible. Herein, the basic principles of molecular cytogenetics, the technologies used to identify chromosomal rearrangements and copy number changes, and the applications for cytogenetics in biomedical diagnosis and research are presented and discussed.

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“…More advanced technologies like the Real-time PCR detection (Ivnitski et al, 1999), DNA extraction, PCR variants and amplicons analysis, Loop mediated isothermal amplification, Nucleic acid sequence based amplification, Biosensors, Electrochemical biosensors (Mortari and Lorenzelli, 2014), Flow cytometry (Shibajyoti Ghosh , Impedance microbiology (S. Ghosh Dweik et al, 2012;Ghosh Dastider et al, 2012) and spectrofluorometry have been employed to detect bacteria (Peacock and Kudela, 2012). Fluorescence in situ hybridization (FISH) is a recent method that represents a rapid and highly specific technique for whole-cell detection (McNamara et al, 2014). However, lack of a protocol for quick detection could lead to significant infection and outbreaks that could rapidly take the form an epidemic, as was the case with the recent outbreak of EHEC O104:H4 infection (http://www.cdc.gov/ecoli/2011/ecolio104/,http://www.euro.who.int/en/healthtopics/emergencies/international-health-regulations/outbreaks-of-e.-coli-o104h4infection).The use of magnetic beads with PCR has also been studied earlier (Fu et al, 2005;Wang et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Highly specific and rapid immuno-fluorescent visualization and detection of E. coli O104:H4 with protein-A coated magnetic beads based LST-MUG assay

Barizuddin

Balakrishnan

Stringer

et al. 2015

Journal of Microbiological Methods

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A method combining immunomagnetic separation and fluorescent sensing was developed to detect Escherichia coli (E. coli) O104:H4. The antibody specific to E. coli O104:H4 was immobilized on protein A-coated magnetic beads. This protein-A-anti E. coli O104:H4 complex was used to bind Fluorescein IsoThioCyanate (FITC) labeled E. coli O104:H4 antigen (whole cell) on it. The goal was to achieve a fluorescently detectable protein-A-anti E. coli O104:H4-E. coli O104:H4 complex on the magnetic beads. Fluorescent microscopy was used to image the magnetic beads. The resulting fluorescence on the beads was due to the FITC labeled antigen binding on the protein-A-anti E. coli O104:H4 immobilized magnetic beads. This visually proves the antigen-antibody binding. The fluorescent imaging results were obtained in 2 h if the minimum available bacteria in the sample were at least 10(5) CFU/ml. If no fluorescence was observed on the magnetic beads during fluorescent imaging, it indicates the bacterial concentration in the sample to be too low for it to have bound to the magnetic beads and hence no detection was possible. To detect bacterial concentration less than 10(5) CFU/ml in the sample, an additional step was required for detection. The magnetic bead complex was added to the LST-MUG (lauryl sulfate tryptose-4-methylumbelliferyl-β-D-glucuronide), a signaling reporter. The E. coli O104:H4 grows in LST-MUG and releases β-glucuronidase enzyme. This enzyme cleaves the MUG substrate that produces 4-methylumbelliferone, a highly fluorescent species. This fluorescence was detected using a spectrofluorometer. The emission peak in the fluorescent spectrum was found to be at 450 nm. The lower and upper detection range for this LST-MUG assay was found to be 2.05×10(5)-4.09×10(8) CFU/ml. The results for the LST-MUG assay for concentrations below 10(5) CFU/ml were ascertained in 8h. The advantages of this technique include the specific detection of bacteria without an enrichment step and allowing the procedure to be completed in hours rather than days.

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The Use of Microarrays and Fluorescence In Situ Hybridization for the Study of Mechanotransduction from Topography

Cited by 6 publications

References 22 publications

Cellular behavior as a dynamic field for exploring bone bioengineering: A closer look at cell–biomaterial interface

Cellular behavior as a dynamic field for exploring bone bioengineering: A closer look at cell–biomaterial interface

Human molecular cytogenetics: from cells to nucleotides

Highly specific and rapid immuno-fluorescent visualization and detection of E. coli O104:H4 with protein-A coated magnetic beads based LST-MUG assay

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