Whole animal images were acquired on an Olympus MVX10 Fluorescence MacroZoom dissecting microscope, using a 540-580 nm excitation filter and 590-670 nm emission filter. Mitochondrial images were taken on a FV1000 Olympus laser scanning confocal microscope using a 100x oil objective (Olympus, N.A. 1.40). Diode laser illumination was 561 nm for red fluorescent transgene and 488 nm for fluorescence of MitoTracker Green FM. Animals were stained with 12 μM MitoTracker Green FM for 20 hr where indicated. MitoTracker stain was dissolved in DMSO, diluted in M9 media (22 mM KH2PO4, 42 mM Na2HPO4, 86 mM NaCl, 1 mM MgSO4, pH 7) and added to OP50 food on culture plates (DMSO <0.02% final concentration) and allowed to dry (Dingley et al. 2010 (link)). Profile plots of pixel intensity were generated using ImageJ software.
Mvx10 fluorescence macrozoom dissecting microscope
Manufactured by Olympus
The MVX10 Fluorescence MacroZoom dissecting microscope is a stereo microscope designed for fluorescence imaging and macro-scale observations. It provides a wide zoom range, high-quality optics, and the capability to visualize fluorescent samples. The microscope is intended for general laboratory and research applications that require observation and analysis of larger specimens.
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3 protocols using mvx10 fluorescence macrozoom dissecting microscope
1
Visualizing Mitochondria in C. elegans
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Mitochondrial Dynamics and Worm Locomotion
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Synchronized day 1 and day 2 adult worms (+/−ATR) were transferred to the freshly seeded plate, and the number of body bends per 15 s was counted. One plate from each group (+/− ATR) was exposed to default white microscopic light, and the rest of the worms were additionally treated with mtOFF-inducing light (0.265 mW/mm2, 540- to 580-nm excitation filter MVX10 Fluorescence MacroZoom dissecting microscope by Olympus powered by an X-Cite 220 V mercury bulb by Excelitas). To measure locomotion upon drug exposure, L4 worms were transferred 24 hours before the experiment to seeded NGM plates containing 10 μM FCCP or 100 μM mdivi-1. For AICAR exposure, day 1 adults were moved to the seeded NGM plate containing 1 mM AICAR 4 hours before the experiment.
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Illumination Sources for Microscopy
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Illumination sources were a 580 nm Quantum SpectraLife LED Hybrid lamp by Quantum Devices, Barneveld WI (abbreviated Quantum LED in the text), a 540‐600 nm GYX module, X‐Cite LED1 by Excelitas, Waltham MA, (abbreviated XCite LED), and a 540‐580 nm excitation filter MVX10 Fluorescence MacroZoom dissecting microscope by Olympus (abbreviated MVX) powered by an X‐Cite 220 V mercury bulb by Excelitas. Light intensities are indicated for each experimental condition and were determined with a calibrated thermopile detector (818P‐010‐12, Newport Corporation, Irvine, CA) and optical power meter (1916‐R, Newport Corporation).
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