Bright field microscopy

Manufactured by Leica

Sourced in Germany, United States

Bright-field microscopy is a type of optical microscopy that uses transmitted light to illuminate the specimen. It is a fundamental technique in the observation and analysis of samples, allowing for the visualization of cellular structures and other microscopic features. The core function of bright-field microscopy is to provide a clear, high-contrast image of the specimen by controlling the illumination and effectively utilizing the differences in refractive index between the sample and the surrounding medium.

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Lab products found in correlation

Horseradish peroxidase detection kit, by Agilent Technologies (3 mentions) Mowiol, by Merck Group (3 mentions) Mab318, by Merck Group (2 mentions) 3 3 diaminobenzidine dab staining, by Vector Laboratories (2 mentions) Glutaraldehyde, by Merck Group (2 mentions) Mgcl2, by Merck Group (2 mentions) Dmlb fluorescence microscope, by Leica (1 mentions) Hematoxylin, by BioVitrum (1 mentions) Cytoselect 24 well wound healing assay, by Cell Biolabs (1 mentions) Vectastain abc kit, by Vector Laboratories (1 mentions) Apoptag peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions) Citric acid buffer, by Vector Laboratories (1 mentions) Animal free blocker, by Vector Laboratories (1 mentions) Anti his tag primary antibody, by Abcam (1 mentions) Quickblock blocking buffer, by Beyotime (1 mentions) Anti gpx4, by Proteintech (1 mentions) Anti steap3, by Proteintech (1 mentions) Dab detection kit, by ZSGB-BIO (1 mentions) Matrigel, by BD (1 mentions) Crystal violet, by Beyotime (1 mentions)

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Fluorescent or bright-field imaging microscopy (4 citations)

38 protocols using bright field microscopy

Quantifying Aortic Atherosclerosis and Hepatic Lipids

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Heart tissues containing the aortic arch were excised from the proximal aortic root to the iliac artery branch and placed in 4% paraformaldehyde environment for 6 hours. The atherosclerotic lesion area of aorta was evaluated by Oil red O staining. Measurement of the lesion size was based on HE staining of paraffin embedded aortic sections. The area and size of the lesion were analyzed by bright-field microscopy (Leica, Wetzlar, Germany) using Image J soft.
Mouse liver tissues were immediately snap-frozen in liquid nitrogen and placed in OCT cryostat embedding compound (Tissue-Tek, Torrance, CA, USA). Frozen liver sections (8 μm) were stained with oil red O according to previous report (18 (link)), and the intracellular lipid droplets were observed and assessed by bright-field microscopy (Leica, Wetzlar, Germany).

Ma C.Y., Shi X.Y., Wu Y.R., Zhang Y., Yao Y.H., Qu H.L., Zhang W., Guo Y.L., Xu R.X, & Li J.J. (2021). Berberine attenuates atherosclerotic lesions and hepatic steatosis in ApoE-/- mice by down-regulating PCSK9 via ERK1/2 pathway. Annals of Translational Medicine, 9(20), 1517.

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Immunohistochemical Analysis of TSPO Expression

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Whole brains were harvested and fixed in 4% buffered formalin for 48 hours, followed by paraffin embedding for histology and immunohistochemistry (IHC). Tissue sections (5.0 μm) were stained with TSPO-specific rabbit polyclonal anti-rat/anti-mouse antibody (Novus Biologicals, LLC, Littleton, CO). Immunoreactivity was assessed using a horseradish peroxidase detection kit (Dako, Glostrup, DK). Hematoxylin and eosin staining was used to evaluate cell density and tumor localization. Sections were visualized and documented using bright field microscopy (Leica Microsystems, Inc., Buffalo Grove, IL, USA).

Tang D., Nickels M.L., Tantawy M.N., Buck J.R, & Manning H.C. (2014). Preclinical imaging evaluation of novel TSPO-PET ligand 2-(5,7-diethyl-2-(4-(2-[18F]fluoroethoxy)phenyl)pyrazolo[1,5-a]pyrimidin-3-yl)-N,N-diethylacetamide ([18F]VUIIS1008) in glioma. Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging, 16(6), 813-820.

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Hydrogel Implant Biocompatibility Assessment

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To assess the integrity and immunogenicity of the hydrogel in vivo (and therefore its functionality as an implant cover), tissue organization was analyzed in paraffin−embedded sections of the trout adipose fin. For this, adipose fin samples fixed in 10% neutral formalin were stored for about 2 months, and then the fixative was replaced with 70% ethanol. Paraffin embedding was carried out according to the processing protocol for biopsy specimens (Table S2). Paraffin molds of dehydrated and paraffin infiltrated tissues were serially sectioned at 6 μm thickness with an RWD Minux S700 microtome (RWD Life Science, Shenzhen, China). Five series of sections with intervals of 40 microns were taken from each sample. The obtained slides were examined using a Leica DMLB fluorescence microscope (Leica Microsystems, Wetzlar, Germany) with a ToupCam UCMOS05100KPA camera (ToupTEK Photonics, Hangzhou, China) to detect fluorescence of dye−loaded microcapsules on histological sections. Next, microsections were deparaffinized and stained with hematoxylin (BioVitrum, St. Petersburg, Russia) and eosin (BioVitrum, Russia) with standard staining procedures. The resulting slides were examined by bright field microscopy (Leica Microsystems, Wetzlar, Germany) to investigate morphological changes at the injection site at the tissue and cellular level.

Borvinskaya E., Matrosova S., Sukhovskaya I., Drozdova P., Titov E., Anikienko I., Lubyaga Y., Gurkov A, & Timofeyev M. (2023). Tissue Reaction to Low-Density Polyacrylamide Gel as a Carrier for Microimplants in the Adipose Fin of Rainbow Trout. Gels, 9(8), 629.

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Evaluating V-AgNPs Efficacy on A549 Spheroids

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To assess efficacy of V-AgNPs on matured 5-days old spheroids, we first prepared A549 spheroids as per the protocol given in method 2.2.4. 5-days old spheroids were then incubated with various concentrations of V-AgNPs that were below LD50-3D dose for 3 days. The images of untreated and V-AgNPs treated spheroids were captured before and after treatment using brightfield microscopy (Leica Microsystems, Germany). From the images, the diameters of A549 spheroids (on 5th day and 8th day) were measured using FIJI software (Fiji Is Just ImageJ, Version 1.53t, National Institutes of Health, USA, Java 1.8.0_322 (64 bit)). The results were represented in terms of mean ± standard error of diameters before and after treatment of V-AgNPs (n = 24).

Joshi A.S., Bapat M.V., Singh P, & Mijakovic I. (2024). Viridibacillus culture derived silver nanoparticles exert potent anticancer action in 2D and 3D models of lung cancer via mitochondrial depolarization-mediated apoptosis. Materials Today Bio, 25, 100997.

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Wound Healing Assay for IPF Fibroblasts

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To assess the effect of esomeprazole and its combination with pirfenidone on the expansion and migration of IPF lung fibroblasts in response to TGFβ, the CytoSelect™ 24-Well Wound Healing Assay (Cell Biolabs; cat # CBA-120) was used. First, the provided inserts were placed inside the wells and 1 × 10⁶ cells were seeded in 250 µL fully-supplemented DMEM. The cells were allowed to form a monolayer around the insert for 24 h. The next day, the cells were synchronized by serum starvation prior to removing the inserts to create a 0.9 mm × 1.8 mm scratch area in each of the wells. The cells were then washed with PBS and imaged for baseline scratch area measurement prior to treatment with vehicle, esomeprazole (100 µM), pirfenidone (1 mM) or the combination for up to 72 h in the absence or presence of TGFβ (10 ng/mL) in triplicates. Finally, images were captured using bright-field microscopy (Leica Microsystems, Germany) to determine the effect of esomeprazole and/or pirfenidone on the migration of IPF lung fibroblasts in response to the mitogenic cytokine TGFβ. The remaining scratch area after treatment was measured using a scaled ruler and was converted into percentage of scratch closure for comparison.

Ebrahimpour A., Ahir M., Wang M., Jegga A.G., Bonnen M.D., Eissa N.T., Montesi S.B., Raghu G, & Ghebre Y.T. (2022). Combination of esomeprazole and pirfenidone enhances antifibrotic efficacy in vitro and in a mouse model of TGFβ-induced lung fibrosis. Scientific Reports, 12, 20668.

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Histological Assessment of Duodenal, Renal, and Hepatic Morphology

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Duodena, kidneys and livers were collected at sacrifice, fixed in 100% ethanol and embedded in paraffin. We collected 5-μm-thick sections using a rotary microtome. We stained the sections with hematoxylin and eosin (H&E) to determine duodenal, renal and hepatic morphology, picrosirius red (PSR) to determine renal fibrosis, and Perls’ prussian blue for iron content. Images were acquired using bright field microscopy (Leica Microsystems, Buffalo Grove, IL, USA.

Francis C., Courbon G., Gerber C., Neuburg S., Wang X., Dussold C., Capella M., Qi L., Isakova T., Mehta R., Martin A., Wolf M, & David V. (2019). Ferric citrate reduces fibroblast growth factor 23 levels and improves renal and cardiac function in a mouse model of chronic kidney disease. Kidney international, 96(6), 1346-1358.

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Quantifying Intervertebral Disc Degeneration

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After MRI scanning, the fixed specimens were decalcified, embedded in resin, and sectioned sagittally at 5 μm intervals. The midsagittal section with the EP injury was then identified. After de-plasticization and re-hydration, the sections were stained with Safranin-O/fastgreen/hematoxylin (SO/FG/H) for disc morphology, glycosaminoglycan (GAG) content, and IVD cellularity. The stained sections were imaged using bright field microscopy (Leica Microsystems, Inc, Deerfield, IL, USA). The severity of IVD degeneration was quantified using a grading system that evaluated NP morphology, NP cellularity, NP-AF border, AF morphology, and EP irregularity [26 (link)] (Figure 9). All spine images were evaluated by three evaluators who were blinded to the experimental groups. The degeneration scores from the three evaluators were averaged for statistical analysis.

Lai A., Iliff D., Zaheer K., Wang D., Gansau J., Laudier D.M., Zachariou V, & Iatridis J.C. (2023). Spinal Cord Sensitization and Spinal Inflammation from an In Vivo Rat Endplate Injury Associated with Painful Intervertebral Disc Degeneration. International Journal of Molecular Sciences, 24(4), 3425.

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Intervertebral Disc Degeneration Scoring

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The formalin-fixed spine specimens were decalcified in formic acid over 3 days with 3 changes, embedded in paraffin, and sectioned sagittally at 5 μm intervals. The mid-sagittal sections with the AF puncture injury were identified and stained with safranin-O/fast-green/hematoxylin for visualizing GAG content, IVD morphology, and IVD cellularity. The stained slides were imaged using bright-field microscopy (Leica Microsystems, Inc., Deerfield, IL, USA). The IVD degeneration score was determined using a semi-quantitative grading system to evaluate NP morphology, NP cellularity, NP-AF border, AF morphology, and EP irregularity [65 (link)]. IVD degeneration grading was performed by an experienced spine researcher who was blinded to the experimental groups. The degeneration scores from the three injured IVDs within the same animal were averaged, and then compared with those isolated from naïve and other AF injury groups.

Lai A., Iliff D., Zaheer K., Gansau J., Laudier D.M., Zachariou V, & Iatridis J.C. (2024). Annulus Fibrosus Injury Induces Acute Neuroinflammation and Chronic Glial Response in Dorsal Root Ganglion and Spinal Cord—An In Vivo Rat Discogenic Pain Model. International Journal of Molecular Sciences, 25(3), 1762.

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TSPO-based Immunohistochemistry of Whole Brains

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Whole brains were harvested and fixed in 4 % buffered formalin for 48 h, followed by paraffin embedding for histology and immunohistochemistry (IHC). Tissue sections (5.0 µm) were stained with TSPO-specific rabbit polyclonal anti-rat/anti-mouse antibody (Novus Biologicals, LLC; Littleton, CO). Immunoreactivity was assessed using a horseradish peroxidase detection kit (Dako, Glostrup, Denmark). Hematoxylin and eosin (H&E) staining was used to evaluate cell density and tumor localization. Sections were visualized and documented using bright field microscopy (Leica Microsystems, Inc.; Buffalo Grove, IL).

Tang D., Li J., Buck J.R., Tantawy M.N., Xia Y., Harp J.M., Nickels M.L., Meiler J, & Manning H.C. (2017). Evaluation of TSPO PET Ligands [18F]VUIIS1009A and [18F]VUIIS1009B: Tracers for Cancer Imaging. Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging, 19(4), 578-588.

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Evaluating Osteocyte Apoptosis in Bone

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We used longitudinal tibia sections for TUNEL and immunostaining. We deparaffinized, rehydrated and incubated the sections in citric acid buffer (10 mmol·L^–1, pH 3) for 60 min at 37 °C (Vector Labs, Burlingame, CA) for antigen retrieval, and 20 min in 1X animal-free blocker (Vector Labs, Burlingame, CA) prior to specific stainings. For detection of endogenous DMP1 in cortical bone, we incubated the sections with anti-DMP1 primary antibody (#ab103203, C-terminal region, Abcam, Cambridge, MA, USA) for 1 h. For detection of injected His-tagged recombinant DMP1 in cortical bone, we incubated sections with anti-His tag primary antibody (Abcam, Cambridge, MA) for 1 h. We then used the immunohistological Vectastain ABC kit (Vector Labs, Burlingame, CA) and performed detection by bright-field microscopy (Leica Microsystems, Buffalo Grove, IL, USA). We performed TUNEL staining using ApopTag Peroxidase In Situ Apoptosis Detection Kit according to manufacturer’s protocol (Millipore Corporation, Temecula, CA) and quantified the ratio of TUNEL-positive osteocytes to total osteocytes on three separate sections per animal.

Dussold C., Gerber C., White S., Wang X., Qi L., Francis C., Capella M., Courbon G., Wang J., Li C., Feng J.Q., Isakova T., Wolf M., David V, & Martin A. (2019). DMP1 prevents osteocyte alterations, FGF23 elevation and left ventricular hypertrophy in mice with chronic kidney disease. Bone Research, 7, 12.

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