Immunofluorescence Staining of Paraffin Sections Step by Step

Zaqout, Sami; Becker, Lena‐Luise; Kaindl, Angela M.

doi:10.3389/fnana.2020.582218

Cited by 121 publications

(61 citation statements)

References 49 publications

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“…The immunofluorescence staining protocol followed was according to the study of Zaquot and colleagues [ 55 ] with minor modifications. All tissues slices were deparaffinized using xylene, ethanol (100%, 95%, 90%, 80%, and 70%), and PBS.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of the Performance of a ZnO-Nanoparticle-Coated Hydrocolloid Patch in Wound Healing

Trinh

Chien

et al. 2022

Polymers

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Hydrocolloid dressings are an important method for accelerating wound healing. A combination of a hydrocolloid and nanoparticles (NPs), such as gold (Au), improves the wound healing rate, but Au-NPs are expensive and unable to block ultraviolet (UV) light. Herein, we combined zinc oxide nanoparticles (ZnO-NPs) with hydrocolloids for a less expensive and more effective UV-blocking treatment of wounds. Using Sprague–Dawley rat models, we showed that, during 10-day treatment, a hydrocolloid patch covered with ZnO-NPs (ZnO-NPs-HC) macroscopically and microscopically stimulated the wound healing rate and improved wound healing in the inflammation phase as shown by reducing of pro-inflammatory cytokines (CD68, IL-8, TNF-α, MCP-1, IL-6, IL-1β, and M1) up to 50%. The results from the in vitro models (RAW264.7 cells) also supported these in vivo results: ZnO-NPs-HCs improved wound healing in the inflammation phase by expressing a similar level of pro-inflammatory mediators (TNF-α and IL-6) as the negative control group. ZnO-NPs-HCs also encouraged the proliferation phase of the healing process, which was displayed by increasing expression of fibroblast biomarkers (α-SMA, TGF-β3, vimentin, collagen, and M2) up to 60%. This study provides a comprehensive analysis of wound healing by measuring the biomarkers in each phase and suggests a cheaper method for wound dressing.

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Section: Methodsmentioning

confidence: 99%

Evaluation of the Performance of a ZnO-Nanoparticle-Coated Hydrocolloid Patch in Wound Healing

Trinh

Chien

et al. 2022

Polymers

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“…The primer sequences are listed in Table 3 . Immunofluorescence (IF) staining of mouse colon sections was performed according to the previously described protocol with slight modifications ( 23 ). Paraffin embedded slides were dewaxed by serial xylene, xylene/ethanol (1:1), 100% ethanol, 95% ethanol, 70% ethanol, and H 2 O.…”

Section: Methodsmentioning

confidence: 99%

Butanol Purified Food Allergy Herbal Formula-2 Has an Immunomodulating Effect ex-vivo in Pediatric Crohn's Disease Subjects

et al. 2021

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Background: TNF-α has a major role in the pathogenesis of Crohn's disease (CD). In contrast, GM-CSF may be beneficial for its anti-inflammatory role in a subset of patients with CD with antibodies against GM-CSF as seen in prior trials of GM-CSF which resulted in clinical improvement in CD. We developed butanol purified Food Allergy Herbal Formula-2 (B-FAHF-2) by refining FAHF-2. FAHF-2 suppressed TNF-α production by human peripheral blood mononuclear cells (PBMCs) and colonic mucosa, and abrogated colitis in a murine model. We sought to examine the effect of B-FAHF-2 and the herbs that comprise it on TNF-α and GM-CSF production as a potential herbal therapy for the treatment of CD.Methods: B-FAHF-2 was examined using high pressure liquid chromatography (HPLC) and compared to the original formulation, FAHF-2. PBMCs from pediatric patients with CD were cultured with lipopolysaccharide and B-FAHF-2, individual herbs or medium alone. Colonic biopsy specimens were cultured with or without B-FAHF-2. TNF-α and GM-CSF were measured by enzyme-linked immunosorbent assay (ELISA). B-FAHF-2 efficacy was tested in vivo in the CD45Rbhi transfer model.Results: B-FAHF-2 had a similar HPLC fingerprint as FAHF-2 but decreased TNF-α production by PBMCs and colonic mucosa from pediatric CD subjects at 20% of the FAHF-2 dose. B-FAHF-2 increased GM-CSF production by PBMCs and colonic mucosa from pediatric CD subjects including those with antibodies to GM-CSF. Of B-FAHF-2's herbal constituents, only Huang Bai suppressed TNF-α and increased GM-CSF production. In the murine model, B-FAHF-2 treatment alleviated colitis.Conclusions: B-FAHF-2 decreased TNF-α production by PBMCs and colonic mucosa from pediatric subjects at a lower dose than FAHF-2. B-FAHF-2 also increased GM-CSF production by PBMCs independent of antibodies. B-FAHF-2 may have a benefit in CD patients.

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“…After they were unfolded, used lter paper to absorb excessive moisture. And put the glass slides into the constant temperature drying oven to dry at 38 ℃ for 6 ~ 8 h. 10min), and set aside [30,31] .…”

Section: Microscopical Identi Cation Of Medicinal Materialsmentioning

confidence: 99%

Pharmacognostic Identification of the Green Peel of Juglans mandshurica Maxim. from Different Regions and Harvest Times

Jielin

Wang

Yan

et al. 2021

Preprint

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Background: The green peel of Juglans mandshurica Maxim. (QLY) can be used as medicine, also known as Qinglongyi[1]. The aim of this study was to identify QLY from the different regions and harvest times by pharmacognostic identification.Methods: In this study, the morphological character, microscopic character and juglone’s content of QLY from different regions and harvest times were compared by morphological identification, paraffin section and powder section, physico-chemical identification and High Performance Liquid Chromatography (HPLC).Results: Morphological identification results that the colour of outer surfaces of peels which harvested from Tangyuan county of Heilongjiang Province in July, August and September are respectively yellow brown, tan and black brown , and the color of the solution graded gradually from light brown yellow to brownish black by water-based test (WBT). The colour of outer surfaces of peels which harvested from Changbai county of Jilin Province in July, August and September are respectively yellowish white, brownish yellow and pale brown, and the color of the solution gradually changed from light brown to dark brown by WBT. As well as the colour of outer surfaces of peels which harvested from Qingyuan county of Liaoning Province in July, August and respectively September are Light yellowish green, light brownish green and brown green, and the color of the solution gradually changed from light green to dark brown by WBT. With the increase of growth period, the number of microscopic characteristics of different Qinglongyi increased. Through physico-chemical identification, it was found that the yield of juglone in QLY picked on September 21, 2016 was the highest, and the content of juglone, whose order was Heilongjiang > Jilin > Liaoning, was as high as 203.476 μg/g.Conclusions: Through comparison, it is concluded that Heilongjiang Province is one of the high-quality producing areas of QLY, and its best harvest time is in the middle and first ten-day period of September to harvest QLY without softening, yellowing and decay. This experiment provides a theoretical basis for the determination of the best harvest conditions in QLY and the establishment of identification standards for medicinal materials.

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Immunofluorescence Staining of Paraffin Sections Step by Step

Cited by 121 publications

References 49 publications

Evaluation of the Performance of a ZnO-Nanoparticle-Coated Hydrocolloid Patch in Wound Healing

Evaluation of the Performance of a ZnO-Nanoparticle-Coated Hydrocolloid Patch in Wound Healing

Butanol Purified Food Allergy Herbal Formula-2 Has an Immunomodulating Effect ex-vivo in Pediatric Crohn's Disease Subjects

Pharmacognostic Identification of the Green Peel of Juglans mandshurica Maxim. from Different Regions and Harvest Times

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