All procedures were performed at 45°C on a microscope stage heated with a thermoplate (TP-110R-100; Tokai Hit, Japan). The cell culture was poured into a tunnel chamber assembled by taping a coverslip (Nakane and Nishizaka, 2017 (link)), and both ends of the chamber were sealed with nail polish to keep from drying the sample. The position of the cell was visualized by infrared light from a halogen lamp with a bandpass filter (FBH850/40; Thorlabs) at a fluence rate of 1 μmol m−2 s−1. The cells were subjected to lateral light stimulus by an LED from the right side of the microscope stage at an angle of 5°. White LEDs at 20 and 500 μmol m−2 s−1 were used as moderate and strong light stimuli for phototaxis, respectively. Blue, teal, green, orange, red, and far-red light were applied by a monochromatic LED, M450LP1, M490L4, M530L3, M625L3, and M730L4 (Thorlabs), respectively. The LED light was collimated by the condenser lens and combined by dichroic mirrors (FF470-Di01, FF509-FDi01, FF560-FDi01, FF685-Di02; Semrock) to apply multicoloured light simultaneously. The wavelength of the resultant light was measured by a spectrometer (BIM-6002A, BroLight, China). Light intensity was measured with a power metre (Q82017A; Advantest, Japan).
Fbh850 40
Manufactured by Thorlabs
The FBH850/40 is a fiber-coupled broadband light source that provides illumination in the 850 nm wavelength range. This product is designed to deliver a stable and broadband optical output for a variety of photonics applications.
Automatically generated - may contain errors
Lab products found in correlation
M450lp1, by Thorlabs (2 mentions)
Ff470 di01, by IDEX Corporation (2 mentions)
Ff509 fdi01, by IDEX Corporation (2 mentions)
Ff560 fdi01, by IDEX Corporation (2 mentions)
Tp 110r 100, by Tokai Hit (2 mentions)
Ff685 di02, by IDEX Corporation (2 mentions)
Aca640 90um cmos camera, by Thorlabs (2 mentions)
Mvl50tm23, by Thorlabs (2 mentions)
4 protocols using fbh850 40
1
Microscopic Phototaxis Assay at Optimal Temperature
2
Larval Imaging with Infrared Illumination
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Larvae were recorded from below using a Basler aca640–90um CMOS camera and a 50 mm focal length c-mount lens (MVL50TM23, Thorlabs) used at a greater distance from the sensor for increased magnification. Darkfield infrared illumination was provided by the collimated output of an 850nm fiber coupled infrared led (M850F2, Thorlabs) aimed at an oblique angle from above the larva to penetrate beneath the objective. An 850 nm bandpass filter (FBH850–40, Thorlabs) was placed in front of the lens to attenuate the incoming light from the Ti:Sa laser.
3
Larval Imaging with Infrared Illumination
4
Phototaxis Analysis of Motile Cells
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All procedures were performed at 45 °C on a microscope stage heated with a thermoplate (TP-110R-100; Tokai Hit, Japan). The cell culture was poured into a tunnel chamber assembled by taping a coverslip (7) (link), and both ends of the chamber were sealed with nail polish to keep from drying the sample. The position of the cell was visualized by infrared light from a halogen lamp with a bandpass filter (FBH850/40; Thorlabs) at a fluence rate of 1 μmol m -2 s -1 . The cells were subjected to lateral light stimulus by an LED from the right side of the microscope stage at an angle of 5 degrees. White LEDs at 20 and 500 μmol m -2 s -1 were used as moderate and strong light stimuli for phototaxis, respectively. Blue, teal, green, orange, red, and far-red light were applied by a monochromatic LED, M450LP1, M490L4, M530L3, M625L3, and M730L4 (Thorlabs), respectively. The LED light was collimated by the condenser lens and combined by dichroic mirrors (FF470-Di01, FF509-FDi01, FF560-FDi01, FF685-Di02; Semrock) to apply multicoloured light simultaneously. The wavelength of the resultant light was measured by a spectrometer (BIM-6002A, BroLight, China). Light intensity was measured with a power metre (Q82017A; Advantest, Japan).
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