Recycling and Long-Term Storage of Fluorescence In Situ Hybridization Slides

Wakai, Susumu; Shibuki, Yasuo; Yokozawa, Karin; Nakamura, Shoko; Adegawa, Yuko; Yoshida, Akihiko; Tsuta, Koji; Furuta, Koh

doi:10.1309/ajcpyx1uti7ldauy

Cited by 3 publications

(1 citation statement)

References 15 publications

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“…Coral tissues for histology-based methods are typically stored as fixed tissues embedded in paraffin blocks or sections mounted on microscopy slides and can be stored at room temperature or 4 °C. For immunolocalization and fluorescence in situ hybridization (FISH), these blocks can retain their quality indefinitely, whereas any generated thin sections from these blocks can deteriorate much more quickly ( Wakai et al, 2014 ; Alamri, Nam & Blancato, 2017 ). Thin sections treated with dyes should be stored in the dark, whereas skeletons and tissue blocks are typically not light sensitive.…”

Section: Considerations For Individual Fields Of Studymentioning

confidence: 99%

Unified methods in collecting, preserving, and archiving coral bleaching and restoration specimens to increase sample utility and interdisciplinary collaboration

Thurber

Schmeltzer

Grottoli

et al. 2022

PeerJ

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Coral reefs are declining worldwide primarily because of bleaching and subsequent mortality resulting from thermal stress. Currently, extensive efforts to engage in more holistic research and restoration endeavors have considerably expanded the techniques applied to examine coral samples. Despite such advances, coral bleaching and restoration studies are often conducted within a specific disciplinary focus, where specimens are collected, preserved, and archived in ways that are not always conducive to further downstream analyses by specialists in other disciplines. This approach may prevent the full utilization of unexpended specimens, leading to siloed research, duplicative efforts, unnecessary loss of additional corals to research endeavors, and overall increased costs. A recent US National Science Foundation-sponsored workshop set out to consolidate our collective knowledge across the disciplines of Omics, Physiology, and Microscopy and Imaging regarding the methods used for coral sample collection, preservation, and archiving. Here, we highlight knowledge gaps and propose some simple steps for collecting, preserving, and archiving coral-bleaching specimens that can increase the impact of individual coral bleaching and restoration studies, as well as foster additional analyses and future discoveries through collaboration. Rapid freezing of samples in liquid nitrogen or placing at −80 °C to −20 °C is optimal for most Omics and Physiology studies with a few exceptions; however, freezing samples removes the potential for many Microscopy and Imaging-based analyses due to the alteration of tissue integrity during freezing. For Microscopy and Imaging, samples are best stored in aldehydes. The use of sterile gloves and receptacles during collection supports the downstream analysis of host-associated bacterial and viral communities which are particularly germane to disease and restoration efforts. Across all disciplines, the use of aseptic techniques during collection, preservation, and archiving maximizes the research potential of coral specimens and allows for the greatest number of possible downstream analyses.

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Section: Considerations For Individual Fields Of Studymentioning

confidence: 99%

Unified methods in collecting, preserving, and archiving coral bleaching and restoration specimens to increase sample utility and interdisciplinary collaboration

Thurber

Schmeltzer

Grottoli

et al. 2022

PeerJ

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A new method for real-time evaluation of pepsin digestion of paraffin-embedded tissue sections, prior to fluorescence in situ hybridisation

et al. 2017

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Fluorescence in situ hybridisation (FISH) is a molecular cytogenetic technique, which is regularly applied to formalin-fixed paraffin-embedded (FFPE) tissue sections of a variety of cancers to assess chromosomal aberrations. However, high-quality FISH requires optimal enzymatic digestion, and insufficient digestion is not noted until the hybridisation signals are evaluated in the fluorescence microscope. As a consequence, FISH results may be unreliable, and the experiment might have to be repeated. To solve this problem, we developed a new method for real-time evaluation of enzymatic tissue digestion. Termination of enzyme activity at the proper time facilitates successful hybridisation, and experiments do not have to be repeated. We first performed FISH on 20 FFPE samples, which had been pepsin digested for different times, and this revealed distinct morphological changes within the nucleus and perinuclear space that were detectable by light microscopy. These observations suggested that the presence of intact and clear bare nuclei, surrounded by a translucent perinuclear space, might serve as an indicator of adequate digestion. We developed a protocol for assessment of this indicator, based on morphological features, and applied this to a collection of 400 tissue samples, partly of breast cancer and partly of different types of lymphoma, prior to FISH. The FISH success rate was 99.5% (398/400), which was significantly higher than that of the conventional method. In all successful cases, morphological signs of adequate digestion were paralleled by easily interpretable FISH signals. This new method for the real-time assessment of digestion quality improved the success rate of FISH and in addition was simple and rapid.

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HER2 fluorescent in situ hybridization signal degradation: a 10-year retrospective study

Galibois

Seebocus

et al. 2021

Breast Cancer Res Treat

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Recycling and Long-Term Storage of Fluorescence In Situ Hybridization Slides

Abstract: Our approach for probe removal and recycling allows repeated examination of even a limited number of slides.

Cited by 3 publications

References 15 publications

Unified methods in collecting, preserving, and archiving coral bleaching and restoration specimens to increase sample utility and interdisciplinary collaboration

Unified methods in collecting, preserving, and archiving coral bleaching and restoration specimens to increase sample utility and interdisciplinary collaboration

A new method for real-time evaluation of pepsin digestion of paraffin-embedded tissue sections, prior to fluorescence in situ hybridisation

HER2 fluorescent in situ hybridization signal degradation: a 10-year retrospective study

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