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Rapid Formation of Plasma Protein Corona Critically Affects Nanoparticle Pathophysiology
Published in Lajos P. Balogh, Nano-Enabled Medical Applications, 2020
Stefan Tenzer, Dominic Docter, Jörg Kuharev, Anna Musyanovych, Verena Fetz, Rouven Hecht, Florian Schlenk, Dagmar Fischer, Klytaimnistra Kiouptsi, Christoph Reinhardt, Katharina Landfester, Hansjörg Schild, Michael Maskos, Shirley K. Knauer, Roland H. Stauber
Confocal laser scanning microscopy. This was employed to determine the cellular localization of nanoparticles as described [44]. Images were acquired using Leica LAS AF software on a Leica SP5 II system (Leica). Further experimental details are provided in the Supplementary Information.
Imaging of Intracellular Calcium in Hippocampal Slices: Methods, Limitations, and Achievements
Published in Avital Schurr, Benjamin M. Rigor, BRAIN SLICES in BASIC and CLINICAL RESEARCH, 2020
Menahem Segal, Jonathan M. Auerbach
Confocal laser scanning microscopes provide the ability to scan a 512-pixel line in the field of view at fast rates (up to 1 kHz).14,15 The confocal laser scanning microscope provides other advantages that make it an ideal research tool for the study of neurons in a tissue slice; the laser light that is focused through a pinhole excites the fluorophore primarily in the plane of focus and can provide an optical section through the tissue, avoiding unnecessary photodynamic damage to the rest of the tissue. Images of sharp focus can thus be obtained from regions deep in the tissue.
Routine and Special Techniques in Toxicologic Pathology
Published in Pritam S. Sahota, James A. Popp, Jerry F. Hardisty, Chirukandath Gopinath, Page R. Bouchard, Toxicologic Pathology, 2018
Daniel J. Patrick, Matthew L. Renninger, Peter C. Mann
Confocal microscopy, or confocal laser scanning microscopy, is a type of optical sectioning microscopy that provides high-resolution images. It shares many of the same principles of conventional wide-field fluorescent microscopy except that excitation and detection are both in focus. To achieve this, the excited light comes from a laser beam and is focused on one point in the specimen (the illumination spot, or Airy disk), the return emitted light from that spot is also focused, and a small pinhole over the detector screens out almost all of the undesirable emitted light outside the plane of focus. Use of these two focal points for illumination (excited light) and detection (emitted light) almost completely eliminates background fluorescence, which markedly increases contrast (Conchello and Lichtman 2005). Typically, to create an image, the illumination spot is moved in a raster fashion (like reading a book) over a thin focal plane section of the specimen, and the two-dimensional image is generated by adding all of the information together. Three-dimensional images can be generated by computationally combining the image data from a stack of two-dimensional images. Extremely fine detail of fluorescently labeled structures near or below the limit of resolution can be visualized, such as cytoskeletal microtubules, organelles, inorganic metallic ions, and receptors.
A critical review on the role of nanotheranostics mediated approaches for targeting β amyloid in Alzheimer’s
Published in Journal of Drug Targeting, 2023
Vaibhav Rastogi, Anjali Jain, Prashant Kumar, Pragya Yadav, Mayur Porwal, Shashank Chaturvedi, Phool Chandra, Anurag Verma
Optical imaging uses visible light and the unique characteristics of photons to produce detailed images of organs, tissues, and even smaller structures like cells and molecules inside the body in a non-invasive manner. In comparison to other conventional techniques, this is one of OI’s biggest advantages and what makes it so user-friendly. It is also comparatively a less expensive technique. Intrinsic tissue absorption and scattering provide information about anatomical features during optical imaging, but it is less informative about specific functionalities (such as metabolism, excretion, and secretion) without the use of fluorescent markers [125]. For the optical imaging, development of confocal laser scanning microscopy (CLSM) offers the fluorescence signal’s axial and lateral interference as well as the ability also allows for optical sectioning, which can be used for three-dimensional imaging of thicker samples.
CDK4/6 blockade provides an alternative approach for treatment of mismatch-repair deficient tumors
Published in OncoImmunology, 2022
Inken Salewski, Julia Henne, Leonie Engster, Paula Krone, Bjoern Schneider, Caterina Redwanz, Heiko Lemcke, Larissa Henze, Christian Junghanss, Claudia Maletzki
Cryostat sections of 4 µm were fixed in cold pure methanol for 8 min, air-dried and unspecific binding site blocked (2% BSA, 2 h) followed by staining with Alexa Fluor 488, Alexa Fluor 594 and Alexa Fluor 647 labeled antibodies CD3, CD4, CD8, CD206, F4/80, CD11b, Gr1, PD-L1, PD-1, and Irf5 (Biolegend). Sections were washed and embedded in Roti Mount Fluor Care DAPI (Roth, Karlsruhe). Visualization was performed on a confocal laser scanning microscope (ZEISS Elyra 7 Confocal Laser Microscope, Zeiss, Jena, Germany). The infiltration pattern was quantified. For infiltrating CD3+CD4+ T helper cells and CD3+CD8+ cytotoxic T cells, numbers were counted in 2–3 high power fields (HPFs)/slide. For regulatory granulocytes and tumor-associated macrophages (TAM), the infiltration pattern was semi-quantitatively analyzed using a scoring system. 0 = no; 1 = mild (1–20 cells/HPF); 2 = moderate (21–40 cells/HPF); 3 = strong (>40 cells/HPF).
Novel biomimetic nanostructured lipid carriers for cancer therapy: preparation, characterization, and in vitro/in vivo evaluation
Published in Pharmaceutical Development and Technology, 2021
Jianwen Zhou, Biru Guo, Wenquan Zhu, Xiaoyu Sui, Xiaoxing Ma, Jiayi Qian, Lixin Cao, Cuiyan Han
NCI-H1299 cells in the logarithmic growth phase were inoculated in 6-well plates, specifically designed for laser confocal microscopy, at a density of 1 × 106 cells/well and were incubated overnight at 37 °C in an air incubator. The culture medium was discarded the next day, and an incomplete RPMI 1640 culture medium containing Cou6, Cou6-NLC, and RBCm-Cou6-NLC (with the same Cou6 concentration at 50 ng/mL) was added. After an additional incubation for 2 h, the culture medium was discarded and the cells were rinsed thrice with cold PBS. Nuclei were stained with Hoechst 33258 for 20 min, and paraformaldehyde (4%) was added for 20 min to fix the cells after washing the cells thrice with cold PBS. Images were viewed with a confocal laser scanning microscope (Zeiss Laser Confocal Microscope, ZEISS-LSM 700, Jena, Germany). The same method was applied to S180 and RAW264.7 cells.