Electron Microscopy of Mouse Lungs

Mouse lungs were inflation-fixed with 2% paraformaldehyde [Electron Microscopy Sciences (EMS)], 2% glutaraldehyde (EMS), 0.1% calcium chloride (Fisher Scientific) in 0.1 M sodium cacodylate (EMS) buffer (SCB), pH 7.2, under 25 cm pressure, followed by immersion fixation with fresh fixative at 4 °C overnight. Lung lobes were cut into 1–2 mm blocks and processed for transmission electron microscopy as previously described^{52 (link)}. 90 nm mouse lung sections were viewed, and images were digitally acquired by a Hitachi H-7650 transmission electron microscope (Hitachi High Technologies USA) equipped with a CCD camera (Advanced Microscopy Techniques) at 80 kV.

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Sitaraman S., Na C.L., Yang L., Filuta A., Bridges J.P, & Weaver T.E. (2019). Proteasome dysfunction in alveolar type 2 epithelial cells is associated with acute respiratory distress syndrome. Scientific Reports, 9, 12509.

Publication 2019

paraformaldehyde glutaraldehyde calcium chloride Hitachi H-7650 CCD camera

Buffer Cacodylate Calcium chloride Electron microscopy Fixative Glutaraldehyde Immersion Lung Microscopy Mouse Paraformaldehyde Pressure Sodium Transmission electron microscope

Corresponding Organization :

Other organizations : Cincinnati Children's Hospital Medical Center, National Jewish Health, University of Colorado Denver

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Variable analysis

independent variables

Fixation with 2% paraformaldehyde, 2% glutaraldehyde, and 0.1% calcium chloride in 0.1 M sodium cacodylate buffer (pH 7.2) under 25 cm pressure

dependent variables

Ultrastructural features of mouse lung sections viewed using transmission electron microscopy

control variables

Immersion fixation with fresh fixative at 4 °C overnight
Cutting lung lobes into 1-2 mm blocks
Preparation of 90 nm mouse lung sections
Viewing sections using a Hitachi H-7650 transmission electron microscope at 80 kV

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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