Cells were grown to mid-exponential phase at 30 °C in YPD and pelleted by centrifugation for 1 min at 4000 rpm (1500 × g) then resuspended in PBS. 5 μL cell suspension was combined with 1 μL of 0.1 g/L Calcofluor White stain and applied to a glass microscope slide. Alternatively, overnight cultures were prepared for each isolate and wild-type strain in yeast extract peptone dextrose (YPD) at 30 °C, with rotation, and then subcultured to mid-log phase before washing twice in 1× PBS and staining with FM4−64 (BioTracker 640 Red C2(FM4−64) Synaptic Dye, Millipore Sigma) at 10 µM for 10 min. Cells were visualized using an Olympus IX70 Epifluorescent Microscope fitted with a Hamamatsu C11440 camera and taken with Olympus CellSens v. 3.2 software.
C11440 camera
Manufactured by Olympus
The C11440 camera is a compact, high-performance digital camera designed for laboratory and research applications. It features a 5.0-megapixel CMOS sensor and a fast, high-resolution image acquisition capability. The camera supports various external triggering modes and offers a range of exposure times to accommodate various experimental requirements.
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Lab products found in correlation
2 protocols using c11440 camera
1
Fluorescent Staining of Yeast Cells
2
Cardiac Activity Imaging in Restrained Flies
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Cardiac activity was imaged from semi-restrained flies using a Hamamatsu C11440 camera (ORCA-flash4.0LT) and a 10X/0.30w objective (Olympus). Excitation light was provided by a 530 nm mounted LED from Thorlabs and a T-cube LED drive from Thorlabs was set at 0.5 A to keep constant illumination intensity. A small porexpam ball (4.3 mm diameter) was held by a custom-made holder (the funnel section of a glass Pasteur pipette) underneath the fly at a distance that allowed the fly to catch the ball. The ball could be freely moved by the fly and if released, the fly could catch the ball again. Visual stimuli were presented on a 24-inch monitor (ASUS VG248QE) with a 144 Hz refresh rate and tilted at 45 degrees over the imaging stage. To visualize the presentation of stimuli in the recorded video, we used a programmable microcontroller (Arduino Mega 2000) to trigger the illumination of a custom-made red LED synchronized with the stimulus presentation. To image the fly’s movement, an infrared (850 nm) LED was placed laterally to illuminate the fly. Fly behavior and ball movement was recorded using a USB3 video camera (Flea3 1.3 MP Mono USB3 Vision, Point Grey, Richmond, Canada) with a 700 nm long pass filter placed 40 cm away from the imaging stage. The entire rig was placed inside a light-tight black enclosure.
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