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Идентификация Рикитсии рикетси в формализованных тканях с парафинами по иммунной флюоресценции.

С небольшим модификацией методов пищеварения трипсина Рикетсиа рикеттсии была продемонстрирована конкретно иммунофлюоресцентной пятнистостью в пятнах тканей, покрытых формальным слоем парафина, от человека, резусной обезьяны и подопытной свиньи с пятнами Рокки Маунтин и в инфицированных мембранах из куриного эмбриона. Ткани были вырезаны на 4 микрона и, используя гелтен в качестве клейки для тканей, были обработаны в обычном порядке. После этого секции переваривались в рефрижераторный 0,1% трипсина в течение 16 часов, промывались и пятна специально для R. rickettsii в результате прямой или кос

Совпадение в аннотации оригинала
With slight modification of a trypsin digestion technique, Rickettsia rickettsii were demonstrated specifically by immunofluorescence staining in Formalin-fixed, paraffin-embedded tissue sections from a human, rhesus monkey, and guinea pig with Rocky Mountain spotted fever and in infected membranes from a chicken embryo. Tissues were cut at 4 micron and, usi…

Comparison of the efficacy of peloid therapy and paraffin treatment in carpal tunnel syndrome.

Carpal tunnel syndrome (CTS) is a common neuropathy caused by median nerve compression at the wrist, managed with treatments like splinting, physical therapy, injections, and surgery. This study aimed to compare the effectiveness of paraffin and peloid therapies combined with night splint use in reducing symptom severity, improving functional status, and enhancing electrophysiological findings in patients with mild to moderate CTS.Sixty patients with mild to moderate CTS scheduled for paraffin or peloid therapy with night splint use were included. Symptom severity was assessed using the Visual…

Detection of Viral Particles in Paraffinized Tissues and Cell Culture by Indirect Immunofluorescence.

Immunofluorescence enables detection of proteins in tissue and cell culture. The specific reaction between the complex antigen-antibody confers security to detection. Disclosure of proteins of interest by fluorochrome allows a perception of its spatial distribution and location. Here we describe the technique of indirect immunofluorescence for detection of E protein from yellow fever virus in paraffinized tissue and in cell cultures.

Establishment of a novel protocol for formalin-fixed paraffin-embedded organoids and spheroids.

Three-dimensional (3D) cell culture models such as spheroids and organoids are widely used in the field of experimental biology. To analyze these 3D experimental models, formalin-fixed paraffin-embedded (FFPE) sections are superior to whole-mount imaging for some experimental purposes, such as exploring samples with a depth limitation of primary antibody penetration immunohistochemically. However, tiny 3D cell culture samples are difficult to embed in paraffin and acquire appropriate sections. In this report, we optimized a protocol of paraffin embedding for spheroids and organoids. In additio…

Panfungal PCR on formalin-fixed, paraffin-embedded tissue: to proceed or not proceed?

Performance of panfungal PCR-DNA sequencing assays for diagnosis of invasive fungal disease on formalin-fixed, paraffin-embedded tissue (FFPE) is influenced by many variables. Interpretation of a positive result can be challenging due to the need to differentiate colonisers and contaminants from clinically significant pathogens. We conducted a retrospective audit on FFPE tissue specimens that underwent panfungal PCR from January 2021 to August 2022. Panfungal PCR results from samples where fungal elements were visualised on histopathology were compared with results from samples where no fungal…

Paraffin histology sections and delayed repair in recurrent atypical fibroxanthoma of the eyelid.

We present a report of a recurrent atypical fibroxanthoma (AFX), a tumour known rarely to involve the eyelid. A 61 year-old male was referred after reappearance of a lower eyelid AFX within two months of resection. The neoplastic spindle cells at the periphery of an AFX may be difficult to differentiate from dermal fibroblasts in frozen sections. Paraffin sections provide better histological definition and thus greater accuracy in determining completeness of excision. The patient's recurrent tumour was surgically excised and overnight paraffin sections were used to obtain margin clearance, bef…

Immunofluorescence Detection of Lysyl Oxidase in Formalin-Fixed Paraffin-Embedded Breast Tissue.

Lysyl oxidase (LOX) is a copper-dependent amine oxidase that catalyzes the oxidative crosslinking of collagen and elastin in the extracellular matrix, contributing to tissue stiffening and remodeling in the tumor microenvironment. Dysregulated LOX expression has been mechanistically linked to breast cancer invasion, pre-metastatic niche formation, and poor clinical prognosis. Here we describe a complete protocol for evaluating LOX protein expression in formalin-fixed paraffin-embedded (FFPE) human breast tissue sections, comprising (1) hematoxylin and eosin (H&E) staining for morphological ass…

High-Throughput Proteomic and Phosphoproteomic Analysis of Formalin-Fixed Paraffin-Embedded Tissues.

Formalin-fixed, paraffin-embedded (FFPE) patient tissues are a valuable resource for proteomic studies with the potential to associate derived molecular insights with clinical annotations and outcomes. Here, we present an optimized, partially automated, plate-based workflow for FFPE proteomics combining pathology-guided macrodissection, xylene-free deparaffinization using Adaptive Focused Acoustics sonication for lysis and decrosslinking, optimized suspension trapping digestion and cleanup of peptides, and LC-MS/MS using Exploris 480, Orbitrap Astral, and timsTOF HT instrumentation. The workfl…

Generative and integrative modeling for transcriptomics with formalin fixed paraffin embedded material.

BACKGROUND: Formalin-fixed paraffin embedded (FFPE) samples suffer from the degradation of nucleic acids, a problem that becomes particularly acute with samples stored for extended periods. It remains challenging to profile FFPE using high-throughput sequencing technologies including RNA-sequencing, and the resulting FFPE RNA-seq (fRNA-seq) data has a high rate of transcript dropout, a property shared with single cell RNA-seq. Transcript counts also have high variance and are prone to extreme values, together making downstream analyses extremely challenging. METHODS: We introduce the PaRaffin…

Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain.

Tissue-clearing and labeling techniques have revolutionized brain-wide imaging and analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce HIF-Clear, a novel method for efficiently clearing and labeling centimeter-thick FFPE specimens using elevated temperature and concentrated detergents. HIF-Clear with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in a whole FFPE mouse brain and is able to be used as the evaluation of disease treatment efficiency. HIF-Clear also supports ex…

Proteomics-Based Analysis and Diagnosis of Formalin-Fixed Paraffin-Embedded Amyloidosis Samples.

Amyloidosis is a group of rare pathologies characterized by abnormal folding and deposition of susceptible proteins in tissues and organs. Diagnosis of amyloidosis often relies on immunohistochemistry of formalin-fixed paraffin-embedded (FFPE) patient samples; however, dependency on antibodies for protein staining is one of the major pitfalls of this approach, especially for the detection of rare amyloidosis types. In recent years, mass spectrometry-based proteomics has emerged as a promising alternative for adequate detection and amyloid typing, despite the fact that preparing FFPE samples fo…

Assessment of panfungal PCR performance with formalin-fixed paraffin-embedded tissue specimens†.

UNLABELLED: We reviewed the performance of a panfungal ITS-2 PCR and Sanger sequencing assay performed on 88 FFPE specimens at The Hospital for Sick Children (Toronto, Canada) in 2019. A potential fungal pathogen was identified by ITS PCR in 62.7 and 2.9% of positive and negative direct slide examination of tissue specimens, respectively. ITS amplicons were detected in 87/88 specimens, with 53/88 (60.2%) considered as 'positive-contaminants' and 34/88 (38.6%) as 'positive-potential pathogen' upon sequencing. Potential pathogens included Blastomyces dermatitidis (17.1%), Cryptococcus neoformans

Profilin 3 genetic architecture in glioma formalin fixed paraffin embedded (FFPE) archive.

Pfn3 is an intron-less gene, encoding actin binding protein that affects structure of cytoskeleton. Although, Pfn3 is mentioned in Allen Brain Atlas and in adult and prenatal Human Brain Tissue Gene Expression Profiles dataset, however, no report on brain and/or brain tumor associated Pfn3 nucleotide sequences are available in the databases. Moreover, pfn3 and pfn4 are always considered as testicular specific genes. The current study explored transcriptional expression profile and genetic architecture of pfn3 in a cohort of fifty formalin fixed paraffin embedded (FFPE) human glioma archive tis…

A novel evaluation method for Ki-67 immunostaining in paraffin-embedded tissues.

The Ki-67 labeling index is traditionally used to investigate tumor aggressiveness. However, no diagnostic or prognostic value has been associated to the heterogeneous pattern of nuclear positivity. The aims of this study were to develop a classification for the patterns of Ki-67-positive nuclei; to search scientific evidence for the Ki-67 expression and location throughout the cell cycle; and to develop a protocol to apply the classification of patterns of Ki-67-positive nuclei in squamous epithelium with different proliferative activities. Based on empirical observation of paraffin sections …

Molecular detection of SARS-CoV-2 in formalin-fixed, paraffin-embedded specimens.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China, in December 2019. The virus rapidly spread globally, resulting in a public health crisis including almost 5 million cases and 323,256 deaths as of May 21, 2020. Here, we describe the identification and evaluation of commercially available reagents and assays for the molecular detection of SARS-CoV-2 in infected FFPE cell pellets. We identified a suitable rabbit polyclonal anti-SARS-CoV spike protein antibody and a mouse monoclonal anti-SARS-CoV nucleoca