Effects of Acetone, Methanol, or Paraformaldehyde on Cellular Structure, Visualized by Reflection Contrast Microscopy and Transmission and Scanning Electron Microscopy

Hoetelmans, Rob W.M.; Prins, Frans A.; Velde, I. Cornelese‐ten; Meer, Joke van der; Velde, C.J. van de; Dierendonck, Jan Hein van

doi:10.1097/00129039-200112000-00010

Cited by 62 publications

(61 citation statements)

References 10 publications

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“…3). This results in inadequate cellular preservation 51,55,156 and a possible shift in the intracellular immunoreactivity, as has been reported for some growth factor peptides. 15 There is extensive variability in Ag recognition among tissues fixed in various fixatives and subsequently embedded in paraffin.…”

Section: Other Fixativesmentioning

confidence: 87%

Technical Aspects of Immunohistochemistry

2005

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Abstract.Immunohistochemistry is an integral technique in many veterinary laboratories for diagnostic and research purposes. In the last decade, the ability to detect antigens (Ags) in tissue sections has improved dramatically, mainly by countering the deleterious effects of formaldehyde with antigen retrieval (AR) and increasing sensitivity of the detection systems. In this review, I address these topics and provide an overview of technical aspects of immunohistochemistry, including those related to antibodies (Abs) and Ags, fixation, AR, detection methods, background, and troubleshooting. Microarray technology and the use of rabbit monoclonal Abs in immunohistochemistry are also discussed.

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Section: Other Fixativesmentioning

confidence: 87%

Technical Aspects of Immunohistochemistry

2005

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“…92 Simultaneously, removal of water destabilizes hydrogen bonding in hydrophilic areas. The resulting protein denaturation causes inadequate cellular preservation 142,153,401 and a possible shift in intracellular immunoreactivity as reported for some growth factor peptides and cytokines (Fig. 6).…”

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confidence: 79%

When Tissue Antigens and Antibodies Get Along

2013

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Once focused mainly on the characterization of neoplasms, immunohistochemistry (IHC) today is used in the investigation of a broad range of disease processes with applications in diagnosis, prognostication, therapeutic decisions to tailor treatment to an individual patient, and investigations into the pathogenesis of disease. This review addresses the technical aspects of immunohistochemistry (and, to a lesser extent, immunocytochemistry) with attention to the antigen-antibody reaction, optimal fixation techniques, tissue processing considerations, antigen retrieval methods, detection systems, selection and use of an autostainer, standardization and validation of IHC tests, preparation of proper tissue and reagent controls, tissue microarrays and other high-throughput systems, quality assurance/quality control measures, interpretation of the IHC reaction, and reporting of results. It is now more important than ever, with these sophisticated applications, to standardize the entire IHC process from tissue collection through interpretation and reporting to minimize variability among laboratories and to facilitate quantification and interlaboratory comparison of IHC results.

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“…S6B). Cytosolic and nuclear proteins are known to be extracted by methanol fixation (27), which is necessary for this protocol, as it removes all non-cross-linked lipids. This may explain the reduced fluorescence intensity in these regions and confirms the GO-term analysis of DAG-interacting proteins, which listed surprisingly many cytosolic proteins.…”

Section: Subcellular Localization Of Lipids and Lipid-interacting Promentioning

confidence: 99%

Trifunctional lipid probes for comprehensive studies of single lipid species in living cells

Höglinger

Nadler

Haberkant

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

111

105

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Lipid-mediated signaling events regulate many cellular processes. Investigations of the complex underlying mechanisms are difficult because several different methods need to be used under varying conditions. Here we introduce multifunctional lipid derivatives to study lipid metabolism, lipid−protein interactions, and intracellular lipid localization with a single tool per target lipid. The probes are equipped with two photoreactive groups to allow photoliberation (uncaging) and photo-cross-linking in a sequential manner, as well as a click-handle for subsequent functionalization. We demonstrate the versatility of the design for the signaling lipids sphingosine and diacylglycerol; uncaging of the probe for these two species triggered calcium signaling and intracellular protein translocation events, respectively. We performed proteomic screens to map the lipid-interacting proteome for both lipids. Finally, we visualized a sphingosine transport deficiency in patient-derived Niemann−Pick disease type C fibroblasts by fluorescence as well as correlative light and electron microscopy, pointing toward the diagnostic potential of such tools. We envision that this type of probe will become important for analyzing and ultimately understanding lipid signaling events in a comprehensive manner.

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Effects of Acetone, Methanol, or Paraformaldehyde on Cellular Structure, Visualized by Reflection Contrast Microscopy and Transmission and Scanning Electron Microscopy

Cited by 62 publications

References 10 publications

Technical Aspects of Immunohistochemistry

Technical Aspects of Immunohistochemistry

When Tissue Antigens and Antibodies Get Along

Trifunctional lipid probes for comprehensive studies of single lipid species in living cells

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