wide field axio observer 7 inverted microscope  (Carl Zeiss)

Journal: Antioxidants

Article Title: Use of Optical Redox Imaging to Quantify Alveolar Macrophage Redox State in Infants: Proof of Concept Experiments in a Murine Model and Human Tracheal Aspirates Samples

doi: 10.3390/antiox13050546

Figure Lengend Snippet: H 2 O 2 -induced changes in AM. ( A ) Mitochondria were stained with MitoView green (images were acquired with a Leica STELLARIS confocal microscope). Under normal control conditions, the alveolar macrophage showed a clear well-defined mitochondrial network structure, whereas the same cell after exposure to exogenous H 2 O 2 (0.45 mM) showed a diffused mitochondrial network structure; ( B , C ) are representative overlay images of control (0 mM H 2 O 2 ) and 0.45 mM H 2 O 2 challenge conditions, respectively. Under control conditions (i.e., ( B )), both red and green signals were recorded. Under 0.45 mM H 2 O 2 challenge (i.e., ( C )), both red and green signals were also recorded. The intensities of red and green fluorescence signals were adjusted to the same color ranges for the same channel for both control and 0.45 mM H 2 O 2 challenge conditions. Visually, ( B ) has more red pixels and less green pixels, indicating less lipid peroxidation. ( C ) has more green pixels and less red pixels, indicating more lipid peroxidation. When red and green pixels co-localize, a yellow pixel appears. ( D ) Shows the quantification of the lipid peroxidation effects induced by exogenous 0.45 mM H 2 O 2 where the intensities of green and red fluorescence are read on the left axis and their ratio on the right axis, N = 10 FOVs, and the error bars are standard errors; the experiment was replicated once with similar results. *, p < 0.05, **, p < 0.01.

Article Snippet: Briefly, ORI was carried out with an inverted wide-field Zeiss fluorescence microscope equipped with a temperature chamber set at 37 °C (Axio Observer 7, ZEISS, Oberkochen, Germany) with a Plan-Apochromat 20×/0.8 M27 lens.

Techniques: Staining, Microscopy, Fluorescence