Fluorescence in situ hybridization (FISH): History, limitations and what to expect from micro-scale FISH?

Huber, Deborah; Voithenberg, Lena Voith von; Kaigala, Govind V.

doi:10.1016/j.mne.2018.10.006

Cited by 118 publications

(96 citation statements)

References 107 publications

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“…We considered whether these subtypes may receive either substantially above- or below-average levels of olfactory stimulation in an unblocked OE. To address this, we used 2-color RNA-FISH to semiquantitatively measure ( Huber et al, 2018 ) S100a5 transcript abundance, which increases with neuronal activity ( Bennett et al, 2010 ; Fischl et al, 2014 ; McClintock et al, 2014 ; Serizawa et al, 2006 ) ( Figure 7A ), within specific OSN subtypes on the 2 sides of the OE of UNO-treated mice, normalized to the average for open-side OSNs ( Figure 7B ). As expected, all of the subtypes analyzed exhibited significantly (p < 0.001) reduced S100a5 mRNA levels on the closed side of the OE relative to the open side.…”

Section: Resultsmentioning

confidence: 99%

Olfactory Stimulation Regulates the Birth of Neurons That Express Specific Odorant Receptors

et al. 2020

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SUMMARY In mammals, olfactory sensory neurons (OSNs) are born throughout life, ostensibly solely to replace damaged OSNs. During differentiation, each OSN precursor “chooses,” out of hundreds of possibilities, a single odorant receptor (OR) gene, which defines the identity of the mature OSN. The relative neurogenesis rates of the hundreds of distinct OSN “subtypes” are thought to be constant, as they are determined by a stochastic process in which each OR is chosen with a fixed probability. Here, using histological, single-cell, and targeted affinity purification approaches, we show that closing one nostril in mice selectively reduces the number of newly generated OSNs of specific subtypes. Moreover, these reductions depend on an animal’s age and/or environment. Stimulation-dependent changes in the number of new OSNs are not attributable to altered rates of cell survival but rather production. Our findings indicate that the relative birth rates of distinct OSN subtypes depend on olfactory experience.

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Section: Resultsmentioning

confidence: 99%

Olfactory Stimulation Regulates the Birth of Neurons That Express Specific Odorant Receptors

et al. 2020

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“…Recent advances in fluorescence in situ hybridization (FISH) experiments have provided a variety of techniques which can be used to study the structure, dynamics and origin of certain loci, chromosomal arms and/or specific chromosomes (reviewed in Cui et al, 2016;Bačovský et al, 2018;Huber et al, 2018). Previous cytogenetic studies in Silene species were based mainly on physical mapping of satellite rDNA (Široký et al, 2001), repeats and transposable elements (Cermak et al, 2008;Kejnovsky et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

The Formation of Sex Chromosomes in Silene latifolia and S. dioica Was Accompanied by Multiple Chromosomal Rearrangements

et al. 2020

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The genus Silene includes a plethora of dioecious and gynodioecious species. Two species, Silene latifolia (white campion) and Silene dioica (red campion), are dioecious plants, having heteromorphic sex chromosomes with an XX/XY sex determination system. The X and Y chromosomes differ mainly in size, DNA content and posttranslational histone modifications. Although it is generally assumed that the sex chromosomes evolved from a single pair of autosomes, it is difficult to distinguish the ancestral pair of chromosomes in related gynodioecious and hermaphroditic plants. We designed an oligo painting probe enriched for X-linked scaffolds from currently available genomic data and used this probe on metaphase chromosomes of S. latifolia (2n = 24, XY), S. dioica (2n = 24, XY), and two gynodioecious species, S. vulgaris (2n = 24) and S. maritima (2n = 24). The X chromosome-specific oligo probe produces a signal specifically on the X and Y chromosomes in S. latifolia and S. dioica, mainly in the subtelomeric regions. Surprisingly, in S. vulgaris and S. maritima, the probe hybridized to three pairs of autosomes labeling their p-arms. This distribution suggests that sex chromosome evolution was accompanied by extensive chromosomal rearrangements in studied dioecious plants.

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“…In the following, we review the different microfluidic platforms and approaches for carrying out FISH assays on-chip. We significantly extend on earlier reviews by Kwasny et al on microfluidic FISH for chromosome abnormalities [33], by Sato on microfluidic FISH for analysis of circulating tumor cells [34] and a historic FISH review with microfluidic FISH outlook by Huber et al [35]. Here, we include microfluidic platforms across the full range of samples, from cells to tissue, from mammalian to microbial samples.…”

Section: Introductionmentioning

confidence: 92%

FISH and chips: a review of microfluidic platforms for FISH analysis

Rodriguez-Mateos

Azevedo

Almeida

et al. 2020

Med Microbiol Immunol

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Fluorescence in situ hybridization (FISH) allows visualization of specific nucleic acid sequences within an intact cell or a tissue section. It is based on molecular recognition between a fluorescently labeled probe that penetrates the cell membrane of a fixed but intact sample and hybridizes to a nucleic acid sequence of interest within the cell, rendering a measurable signal. FISH has been applied to, for example, gene mapping, diagnosis of chromosomal aberrations and identification of pathogens in complex samples as well as detailed studies of cellular structure and function. However, FISH protocols are complex, they comprise of many fixation, incubation and washing steps involving a range of solvents and temperatures and are, thus, generally time consuming and labor intensive. The complexity of the process, the relatively high-priced fluorescent probes and the fairly high-end microscopy needed for readout render the whole process costly and have limited wider uptake of this powerful technique. In recent years, there have been attempts to transfer FISH assay protocols onto microfluidic lab-on-a-chip platforms, which reduces the required amount of sample and reagents, shortens incubation times and, thus, time to complete the protocol, and finally has the potential for automating the process. Here, we review the wide variety of approaches for lab-on-chip-based FISH that have been demonstrated at proof-of-concept stage, ranging from FISH analysis of immobilized cell layers, and cells trapped in arrays, to FISH on tissue slices. Some researchers have aimed to develop simple devices that interface with existing equipment and workflows, whilst others have aimed to integrate the entire FISH protocol into a fully autonomous FISH on-chip system. Whilst the technical possibilities for FISH on-chip are clearly demonstrated, only a small number of approaches have so far been converted into off-the-shelf products for wider use beyond the research laboratory. Keywords Microfluidics-assisted FISH • µFISH • Microfluidics • Lab-on-a-Chip (LOC) • Fluorescence in situ hybridization (FISH) Edited by Volkhard A. J. Kempf.

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Fluorescence in situ hybridization (FISH): History, limitations and what to expect from micro-scale FISH?

Cited by 118 publications

References 107 publications

Olfactory Stimulation Regulates the Birth of Neurons That Express Specific Odorant Receptors

Olfactory Stimulation Regulates the Birth of Neurons That Express Specific Odorant Receptors

The Formation of Sex Chromosomes in Silene latifolia and S. dioica Was Accompanied by Multiple Chromosomal Rearrangements

FISH and chips: a review of microfluidic platforms for FISH analysis

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