Proximity probing assays for simultaneous visualization of protein complexesin situ

2013 ◽  
Vol 10 (3) ◽  
pp. 219-221 ◽  
Author(s):  
José Manuel Afonso Moreira ◽  
Stine Buch Thorsen ◽  
Nils Brünner ◽  
Jan Stenvang
Keyword(s):  
Simultaneous Visualization

Three-Dimensional Subcellular Organization of Hepatocytes in Intact Liver: Simultaneous Visualization of Cytoskeletal and Membrane Markers by Confocal Microscopy

1996 ◽  
Vol 54 ◽  
pp. 288-289
Author(s):  
Greg V. Martin ◽  
Ann L. Hubbard
Keyword(s):  
Three Dimensional ◽  
Integral Membrane Proteins ◽  
Polarized Secretion ◽  
Microscope Images ◽  
Computer Aided ◽  
Intracellular Organelles ◽  
Cytoskeletal Elements ◽  
Simultaneous Visualization

The microtubule (MT) cytoskeleton is necessary for many of the polarized functions of hepatocytes. Among the functions dependent on the MT-based cytoskeleton are polarized secretion of proteins, delivery of endocytosed material to lysosomes, and transcytosis of integral plasma membrane (PM) proteins. Although microtubules have been shown to be crucial to the establishment and maintenance of functional and structural polarization in the hepatocyte, little is known about the architecture of the hepatocyte MT cytoskeleton in vivo, particularly with regard to its relationship to PM domains and membranous organelles. Using an in situ extraction technique that preserves both microtubules and cellular membranes, we have developed a protocol for immunofluorescent co-localization of cytoskeletal elements and integral membrane proteins within 20 µm cryosections of fixed rat liver. Computer-aided 3D reconstruction of multi-spectral confocal microscope images was used to visualize the spatial relationships among the MT cytoskeleton, PM domains and intracellular organelles.

scholarly journals Simultaneous Visualization of 161Tb- and 177Lu-Labeled Somatostatin Analogues Using Dual-Isotope SPECT Imaging

Pharmaceutics ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 536
Author(s):  
Francesca Borgna ◽  
Patrick Barritt ◽  
Pascal V. Grundler ◽  
Zeynep Talip ◽  
Susan Cohrs ◽  
...  
Keyword(s):  
Pharmacokinetic Profile ◽  
Somatostatin Analogues ◽  
Distribution Coefficients ◽  
Spect Imaging ◽  
Cell Uptake ◽  
Organ Distribution ◽  
Dual Isotope ◽  
Biodistribution Studies ◽  
Simultaneous Visualization

The decay of terbium-161 results in the emission of β¯-particles as well as conversion and Auger electrons, which makes terbium-161 interesting for therapeutic purposes. The aim of this study was to use dual-isotope SPECT imaging in order to demonstrate visually that terbium-161 and lutetium-177 are interchangeable without compromising the pharmacokinetic profile of the radiopharmaceutical. The 161Tb- and 177Lu-labeled somatostatin (SST) analogues DOTATOC (agonist) and DOTA-LM3 (antagonist) were tested in vitro to demonstrate equal properties regarding distribution coefficients and cell uptake into SST receptor-positive AR42J tumor cells. The radiopeptides were further investigated in AR42J tumor-bearing nude mice using the method of dual-isotope (terbium-161/lutetium-177) SPECT/CT imaging to enable the visualization of their distribution profiles in the same animal. Equal pharmacokinetic profiles were demonstrated for either of the two peptides, irrespective of whether it was labeled with terbium-161 or lutetium-177. Moreover, the visualization of the sub-organ distribution confirmed similar behavior of 161Tb- and 177Lu-labeled SST analogues. The data were verified in quantitative biodistribution studies using either type of peptide labeled with terbium-161 or lutetium-177. While the radionuclide did not have an impact on the organ distribution, this study confirmed previous data of a considerably higher tumor uptake of radiolabeled DOTA-LM3 as compared to the radiolabeled DOTATOC.

Dual scope method: A novel application of a simultaneous multi-image display system for a thoracoscopic robotic lobectomy

2021 ◽  
Keyword(s):  
Thoracic Surgery ◽  
Image Display ◽  
The Novel ◽  
Thoracic Cavity ◽  
Display System ◽  
Operative Field ◽  
Robot Arms ◽  
Simultaneous Visualization

The advantages of a multi-input display system platform in robotic thoracic surgery have not been well described. We report the novel application of a multi-display system for simultaneous visualization of an additional thoracoscopic image during a robotic lobectomy, which we have named the dual scope method. An additional thoracoscope is inserted from the bottom of the thoracic cavity. This thoracoscope visualizes the whole operative field, including the robot arms, from a bystander’s viewpoint. By providing an integrated image from the robot scope and the thoracoscope, various problems, such as arm collision, inappropriate instrument direction, excessive traction, and injury, can be solved or avoided much more easily and safely than with the use of the robotic image alone. The dual scope method facilitates the safety and efficiency of robotic lobectomy.

scholarly journals Combined Autoradiography and Formaldehyde-induced Fluorescence Methods for Localization of Radioactively Labeled Substances in Relation to Monoamine Neurons

1981 ◽  
Vol 29 (1A_suppl) ◽  
pp. 175-180 ◽  
Author(s):  
Lester D. Grant ◽  
Walter E. Stumpf
Keyword(s):  
Brain Regions ◽  
Drug Uptake ◽  
Monoamine Neurons ◽  
Freeze Dried ◽  
Uptake Sites ◽  
Lower Brain Stem ◽  
Processing Steps ◽  
Combined Technique ◽  
Histological Staining ◽  
Simultaneous Visualization

A combined technique of formaldehyde-induced fluorescence (FIF) and autoradiography is described for the localization of radioactively labeled substances in relation to monoamine neurons. This method permits the simultaneous visualization of 3H-labeled steroid hormone or drug uptake sites and fluorescing monoamine neural elements (cell bodies, fiber projections, terminals) in the same tissue section. Thin frozen sections cut in a cryostat are freeze-dried, exposed to formaldehyde vapor at 80°C, and carried through dry-mount autoradiography processing steps before fluorescence microscopy screening. Subsequent histological staining of sections and light microscopy are employed for conventional autoradiogram screening. With this procedure, 3H-estradiol and 3H-dihydro-testosterone are localized in various catecholamine (CA) neurons in the diencephalon and lower brain stem of the rat. Also, catecholaminergic as well as noncatechol-aminergic sex steroid target neurons are seen to be innervated by CA terminals in various rat brain regions.

scholarly journals The ER Stress Surveillance (ERSU) pathway regulates daughter cell ER protein aggregate inheritance

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Francisco J Piña ◽  
Maho Niwa
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Mother Cell ◽  
Daughter Cell ◽  
Protein Aggregates ◽  
Cytoplasmic Protein ◽  
Protein Aggregate ◽  
Daughter Cells ◽  
Simultaneous Visualization

Stress induced by cytoplasmic protein aggregates can have deleterious consequences for the cell, contributing to neurodegeneration and other diseases. Protein aggregates are also formed within the endoplasmic reticulum (ER), although the fate of ER protein aggregates, specifically during cell division, is not well understood. By simultaneous visualization of both the ER itself and ER protein aggregates, we found that ER protein aggregates that induce ER stress are retained in the mother cell by activation of the ER Stress Surveillance (ERSU) pathway, which prevents inheritance of stressed ER. In contrast, under conditions of normal ER inheritance, ER protein aggregates can enter the daughter cell. Thus, whereas cytoplasmic protein aggregates are retained in the mother cell to protect the functional capacity of daughter cells, the fate of ER protein aggregates is determined by whether or not they activate the ERSU pathway to impede transmission of the cortical ER during the cell cycle.

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