1x81 inverted epifluorescence microscope

Manufactured by Olympus

The 1X81 inverted epifluorescence microscope is a laboratory equipment designed for fluorescence microscopy. It features an inverted optical configuration and provides illumination for epifluorescence imaging. The core function of this microscope is to enable the observation and analysis of fluorescently labeled samples.

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Tcs sp5 2, by Leica (1 mentions)

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Inverted microscope (2417 citations) Epifluorescence microscope (122 citations) Inverted epifluorescence microscope (25 citations) 1X81 microscope (10 citations) 1X81 inverted microscope (2 citations)

3 protocols using 1x81 inverted epifluorescence microscope

Quantification of DHE Oxidation Products

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Dihydroethidium (DHE) oxidation products were quantified as described previously (28 ). Briefly, HUVEC or HAEC were incubated with scrambled-tat or gp91ds-tat (30 μM) for 1 h, then stimulated with LPS (200 ng/mL) for 4 h. Cells were washed and incubated with DHE (100 μM, 37°C), then scraped in acetonitrile for lyophilization. Quantification of DHE products was performed by high-performance liquid chromatography (HPLC).
DHE fluorescence was used to estimate ROS production in mesenteric vessels. For this, 7-μm-thick tissue sections were incubated with 3 μM DHE in PBS with DTPA (diethylenetriamine-pentacetic acid 100 μM) for 30 min and then images obtained immediately on an Olympus 1X81 inverted epifluorescence microscope.

Trevelin S.C., Sag C.M., Zhang M., Alves-Filho J.C., Cunha T.M., dos Santos C.X., Sawyer G., Murray T., Brewer A., Laurindo F.R., Protti A., Lopes L.R., Ivetic A., Cunha F.Q, & Shah A.M. (2020). Endothelial Nox2 Limits Systemic Inflammation and Hypotension in Endotoxemia by Controlling Expression of Toll-Like Receptor 4. Shock (Augusta, Ga.), 56(2), 268-277.

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Quantifying Oxidative Stress in Aortic Tissue and Fibroblasts

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Dihydroethidium fluorescence was used to estimate ROS production in aorta. For this, 7-µm thick tissue sections were incubated with 3 µmol/L dihydroethidium in Hank’s without phenol red, with diethylenetriamine-pentaacetic acid (100 µmol/L) for 30 minutes. Images were then obtained immediately on an Olympus 1X81 inverted epifluorescence microscope.
The dihydroethidium oxidation product, 2-hydroxyethidium, was quantified in aorta and cultures of fibroblasts as previously described.^{16 (link)} Aortae from mice chronically infused with ANG II or fibroblasts stimulated with ANG II (200 nmol/L, 4 hours) were incubated with dihydroethidium (100 μM) for 30 minutes at 37 °C and then lyophilized in acetonitrile. Quantification of 2-hydroxyethidium was performed by high-performance liquid chromatography (HPLC).

Harrison C.B., Trevelin S.C., Richards D.A., Santos C.X., Sawyer G., Markovinovic A., Zhang X., Zhang M., Brewer A.C., Yin X., Mayr M, & Shah A.M. (2020). Fibroblast Nox2 (NADPH Oxidase-2) Regulates ANG II (Angiotensin II)–Induced Vascular Remodeling and Hypertension via Paracrine Signaling to Vascular Smooth Muscle Cells. Arteriosclerosis, Thrombosis, and Vascular Biology, 41(2), 698-710.

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Immunofluorescence Staining Protocol

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Cells or 7-μm-thick tissue sections were fixed with 4% paraformaldehyde for 30 min and permeabilized in 0.2% Triton-X for 30 min. Unspecific interactions were blocked by incubation with 2% BSA plus 1:50 goat serum solution for 30 min. Primary antibodies were used at 1:100 to 1:200 dilution. Images were acquired on a confocal microscope (TCS SP5 II, Leica) or an Olympus 1X81 inverted epifluorescence microscope.

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