FISH and chips: Marine bacterial communities analyzed by flow cytometry based on microfluidics

Gerdts, Gunnar; Luedke, Gerd

doi:10.1016/j.mimet.2005.05.001

Cited by 23 publications

(16 citation statements)

References 41 publications

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“…8A) 165–167 . This instrument can perform several routine diagnostics using a variety of microfluidic chips, one of which is a flow cytometry chip that enables direct cell or particle analysis based on fluorescence and scatter properties.…”

Section: Integration Of Microfluidics and Microfabrication Into Flmentioning

confidence: 99%

The intersection of flow cytometry with microfluidics and microfabrication

2014

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A modern flow cytometer can analyze and sort particles on a one by one basis at rates of 50,000 particles per second. Flow cytometers can also measure as many as 17 channels of fluorescence, several angles of scattered light, and other non-optical parameters such as particle impedance. More specialized flow cytometers can provide even greater analysis power, such as single molecule detection, imaging, and full spectral collection, at reduced rates. These capabilities have made flow cytometers an invaluable tool for numerous applications including cellular immunophenotyping, CD4+ T-cell counting, multiplex microsphere analysis, high-throughput screening, and rare cell analysis and sorting. Many bio-analytical techniques have been influenced by the advent of microfluidics as a component in analytical tools and flow cytometry is no exception. Here we detail the functions and uses of a modern flow cytometer, review the recent and historical contributions of microfluidics and microfabricated devices to field of flow cytometry, examine current application areas, and suggest opportunities for the synergistic application of microfabrication approaches to modern flow cytometry.

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Section: Integration Of Microfluidics and Microfabrication Into Flmentioning

confidence: 99%

The intersection of flow cytometry with microfluidics and microfabrication

2014

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“…However, we are aware of relatively few applications of microfluidic technologies to microbial FISH, either mRNA or rRNA. The earliest example we are aware of is the use of the Agilent Bioanalyzer 2100 flow cytometry chip, to perform flow analysis of FISH-labeled marine microbes [6], or FISH-labeled Pseudomonas cells in milk [42]. However, in these examples, FISH hybridization was performed in a conventional protocol using microbes in suspension, or enriched on a filter membrane, followed by resuspension of microbes and introduction into the Bioanalyzer chip.…”

Section: Microfluidic Fish For Microbial Rnamentioning

confidence: 99%

Microfluidic Approaches to Fluorescence In Situ Hybridization (FISH) for Detecting RNA Targets in Single Cells

Meagher

2016

Microfluidic Methods for Molecular Biology

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Fluorescence in situ hybridization (FISH) is a powerful molecular technique in cell biology and microbiology for detection and localization of a nucleic acid target within an intact cell or chromosome spread, based on hybridization of a fluorescently labeled nucleic acid "probe" to its complementary target. In some instances, FISH analysis is performed on intact samples-whether thin tissue sections, or environmental samples, allowing the nucleic acid target to be localized in context with other cells. FISH evolved from in situ hybridization (ISH) techniques utilizing radiolabeled probes. By comparison, FISH typically utilizes smallmolecule fluorophores. This labeling approach eliminates the hazards associated with radioactivity, and allows analysis with common laboratory instrumentation, including epifluorescence or laser-scanning confocal microscopes, or flow cytometers. Unlike PCR, sequencing, or most other nucleic acid analysis methods, FISH is fundamentally a single-cell measurement technique. Whether using imaging or flow cytometry as a readout, the signals from FISH are detected and analyzed on a cell-by-cell basis, affording a unique capability for studying rare events or heterogeneity within a population. Coupling of FISH with flow sorters also affords a unique capability for enriching cell or chromosome populations or even isolating single cells based on presence of specific nucleic acid targets [7,10,43].

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“…Only a few studies (Lebaron et al, 1997;Gerdts & Luedk, 2006;Kalyuzhnaya et al, 2006;Yilmaz et al, 2010) have combined FISH and FCM for the analysis of acquatic microbial communities. The main limitation of combining CARD-FISH and FCM is that the former is commonly performed and optimized on a solid support (i.e.…”

Section: Bacteria Identification With Antibodies and Nucleic Acid Promentioning

confidence: 99%

What Flow Cytometry can Tell Us About Marine Micro-Organisms – Current Status and Future Applications

Manti¹,

Papa²,

Boi³

2012

Flow Cytometry - Recent Perspectives

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FISH and chips: Marine bacterial communities analyzed by flow cytometry based on microfluidics

Cited by 23 publications

References 41 publications

The intersection of flow cytometry with microfluidics and microfabrication

The intersection of flow cytometry with microfluidics and microfabrication

Microfluidic Approaches to Fluorescence In Situ Hybridization (FISH) for Detecting RNA Targets in Single Cells

What Flow Cytometry can Tell Us About Marine Micro-Organisms – Current Status and Future Applications

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