Cubic Clearing and Whole Mount Imaging of Mouse Lung Lobes v1

protocols.io ◽  
2021 ◽  
Author(s):  
Jamie not provided Verheyden
Keyword(s):  
Mouse Lung ◽  
Whole Mount ◽  
Lung Lobes

Protocol for CUBIC Clearing and Whole Mount Imaging of Mouse Lung Lobes v1

2021 ◽  
Author(s):  
Justinn Barr ◽  
Jamie Verheyden ◽  
Xin Sun
Keyword(s):  
Laboratory Animal ◽  
Computational Analysis ◽  
American Association ◽  
Mouse Lung ◽  
Whole Mount ◽  
Mouse Lung Tissue ◽  
Laboratory Animal Facility ◽  
The University ◽  
Animal Care ◽  
Lung Lobes

This protocol is for Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis (CUBIC) of mouse lung tissue for whole lobe imaging using Zeiss Lightsheet Imaging. All experimental procedures were performed in the American Association for Accreditation of Laboratory Animal Care (AAALAC)-certified laboratory animal facility at the University of California San Diego, following protocols approved by the institutional animal care and use committee (IACUC). The procedures should incorporate all local requirements for standards of animal experimentation.

scholarly journals Stereological analysis of individual lung lobes during normal and aberrant mouse lung alveolarisation

2018 ◽  
Vol 232 (3) ◽  
pp. 472-484 ◽  
Author(s):  
Tuong-Van Hoang ◽  
Claudio Nardiello ◽  
David E. Surate Solaligue ◽  
José Alberto Rodríguez-Castillo ◽  
Philipp Rath ◽  
...  
Keyword(s):  
Mouse Lung ◽  
Stereological Analysis ◽  
Lung Lobes

Estimation of 100 nm particle distribution kinetics in mouse lung using confocal laser scanning microscopy

2021 ◽  
Vol 21 (3) ◽  
pp. 103-108
Author(s):  
Julia D. Vavilova ◽  
Elena L. Bolkhovitina ◽  
Andrey O. Bogorodskiy ◽  
Ivan S. Okhrimenko ◽  
Valentin I. Borshchevskiy ◽  
...  
Keyword(s):  
Respiratory Tract ◽  
Laser Scanning ◽  
Particle Distribution ◽  
Laser Scanning Microscopy ◽  
Mouse Lung ◽  
Confocal Laser ◽  
Phagocytic Cells ◽  
Viral Particles ◽  
Mouse Lungs ◽  
Lung Lobes

BACKGROUND: Daily, people inhale airborne viral particles, some of which have a size of about 100 nm, such as particles of SARS-CoV-2. Kinetics of such 100 nm particle distribution in the respiratory tract is important, however, not a properly investigated question. AIM: To estimate the dissemination of inert viral particles based on the analysis of the spatial distribution of fluorescent 100 nm particles in the mouse lungs at different time points after the application. MATHERIALS AND METHODS: Fluorescent particles of 100 nm size were applied to C57BL/6 mice. 6, 24, 48 and 72 hours after, lungs were excised and fixed. Lung lobes were stained with immunohistochemistry as whole-mounts and then underwent optical clearance. Three-dimensional images of whole-mount mouse lung lobes were acquired using confocal laser scanning microscopy. RESULTS: 6 hours after the particle application particles were detected in lungs both as single particles and as particle agglomerates. Particles were both free and internalized by phagocytic cells. 24 hours after the application particles were detected both in bronchial lumen and in the alveolar space. Particles were detected in the mouse lungs up to 72 hours after the application. CONCLUSIONS: Reaching the respiratory tract of mammalian, inert particles which size equal to SARS-CoV-2 particle size distribute both in bronchi and in alveoli and undergoes internalization of phagocytic cells.

Imaging of the mouse lung with scanning laser optical tomography (SLOT)

2012 ◽  
Vol 113 (6) ◽  
pp. 975-983 ◽  
Author(s):  
Manuela Kellner ◽  
Marko Heidrich ◽  
Rebecca Beigel ◽  
Raoul-Amadeus Lorbeer ◽  
Lars Knudsen ◽  
...  
Keyword(s):  
Internal Structure ◽  
Ex Vivo ◽  
Optical Tomography ◽  
Three Dimensional ◽  
Scanning Laser ◽  
Mouse Lung ◽  
Microscopy Technique ◽  
Planar Orientation ◽  
Scan Modes ◽  
Lung Lobes

The current study focuses on the use of scanning laser optical tomography (SLOT) in imaging of the mouse lung ex vivo. SLOT is a highly efficient fluorescence microscopy technique allowing rapid scanning of samples of a size of several millimeters, thus enabling volumetric visualization by using intrinsic contrast mechanisms of previously fixed lung lobes. Here, we demonstrate the imaging of airways, blood vessels, and parenchyma from whole, optically cleared mouse lung lobes with a resolution down to the level of single alveoli using absorption and autofluorescence scan modes. The internal structure of the lung can then be analyzed nondestructively and quantitatively in three-dimensional datasets in any preferred planar orientation. Moreover, the procedure preserves the microscopic structure of the lung and allows for subsequent correlative histologic studies. In summary, the current study has shown that SLOT is a valuable technique to study the internal structure of the mouse lung.

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

1988 ◽  
Vol 46 ◽  
pp. 30-31
Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis
Keyword(s):  
Whole Blood ◽  
Colloidal Gold ◽  
Blood Platelets ◽  
Cell Contact ◽  
Computer Assisted ◽  
Adherent Cells ◽  
Differential Centrifugation ◽  
Computer Reconstruction ◽  
Sds Page ◽  
Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

Applications of Scanning Microscopy in Biomedical Research

1968 ◽  
Vol 26 ◽  
pp. 354-355
Author(s):  
T. L. Hayes
Keyword(s):  
Information Content ◽  
Biomedical Research ◽  
Biomedical Applications ◽  
Three Dimensional ◽  
Dental Enamel ◽  
Resolving Power ◽  
Whole Mount ◽  
Two Parameters ◽  
Content Information ◽  
Sectioned Tissue

Biomedical applications of the scanning electron microscope (SEM) have increased in number quite rapidly over the last several years. Studies have been made of cells, whole mount tissue, sectioned tissue, particles, human chromosomes, microorganisms, dental enamel and skeletal material. Many of the advantages of using this instrument for such investigations come from its ability to produce images that are high in information content. Information about the chemical make-up of the specimen, its electrical properties and its three dimensional architecture all may be represented in such images. Since the biological system is distinctive in its chemistry and often spatially scaled to the resolving power of the SEM, these images are particularly useful in biomedical research.In any form of microscopy there are two parameters that together determine the usefulness of the image. One parameter is the size of the volume being studied or resolving power of the instrument and the other is the amount of information about this volume that is displayed in the image. Both parameters are important in describing the performance of a microscope. The light microscope image, for example, is rich in information content (chemical, spatial, living specimen, etc.) but is very limited in resolving power.

The application of video-enhanced constrast/differential interference constrast microscopy in conjunction with whole mount electron microscopy to the study of organelle transport

1988 ◽  
Vol 46 ◽  
pp. 66-67
Author(s):  
J. H. Hayden
Keyword(s):  
Electron Microscopy ◽  
Rana Pipiens ◽  
Silver Film ◽  
Immunofluorescence Microscopy ◽  
Organelle Transport ◽  
Linear Element ◽  
Whole Mount ◽  
Carbon Coated ◽  
Linear Elements ◽  
Dic Microscopy

In a previous study, Allen video-enhanced constrast/differential interference constrast (AVEC-DIC) microscopy was used in conjunction with immunofluorescence microscopy to demonstrate that organelles and vesicle move in either direction along linear elements composed of microtubules. However, this study was limited in that the number of microtubules making up a linear element could not be determined. To overcome this limitation, we have used AVEC-DIC microscopy in conjunction with whole mount electron microscopy.Keratocytes from Rana pipiens were grown on glass coverslips as described elsewhere. Gold London Finder grids were Formvar- and carbon coated, and sterilized by exposure to ultraviolet light. It is important to select a Formvar film that gives a grey reflection when it is floated on water. A silver film is too thick and will detract from the image in the light microscope.

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