Experiments were performed by EPC 800 patch clamp amplifier (HEKA Electronic GmbH, Pfalz, Germany). QDAS integrated and control devices were cleaned with 70 vol.% ethanol solution and incubated for 3 days in DI water. The pulled patch pipettes of 4–6 m Ω were utilized to carry out the whole‐cell patch clamp experiment. The internal cellular medium was prepared by mixing 140 mm KCl, 2 mm MgCl2, 10 mm HEPES, 10 mm ethylene glycol‐bis (β‐aminoethyl ether)‐N,N,N′,N′‐tetraacetic acid (EGTA), 2 mm Mg‐ATP in water, and the pH was calibrated to 7.2–7.3 using 1 m KOH. The intracellular solution was filled into the patch pipettes to achieve whole‐cell patch. A digital camera integrated Olympus T2 upright microscope was used to patch and monitor the cells. The blue LED (M450LP1, Thorlabs Inc, NJ, USA) was used as the light source. The LED system was driven by DC2200 – High‐Power 1‐ Channel LED Driver (Thorlabs Inc., NJ, USA).
M450lp1
Manufactured by Thorlabs
Sourced in United States
The M450LP1 is a 450 nm longpass filter from Thorlabs. It is designed to transmit light at wavelengths longer than 450 nm while blocking shorter wavelengths.
Automatically generated - may contain errors
Lab products found in correlation
T2 upright microscope, by Olympus (2 mentions)
Dc2200 high power 1 channel led driver, by Thorlabs (2 mentions)
Fbh850 40, 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)
Ff01 535 50, by IDEX Corporation (1 mentions)
Ff495 di03, by IDEX Corporation (1 mentions)
M405lp1, by Thorlabs (1 mentions)
Scmos camera, by Oxford Instruments (1 mentions)
Igor pro, by Wavemetrics (1 mentions)
Pci 6229, by National Instruments (1 mentions)
Ledd1b, by Thorlabs (1 mentions)
Dcc1545m, by Thorlabs (1 mentions)
8 protocols using m450lp1
1
Whole-Cell Patch Clamp Technique
2
Microscopic Phototaxis Assay at Optimal Temperature
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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 (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).
3
High-Resolution Fluorescence Microscopy Imaging
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Images of HEK293 cells or neurons were acquired with an inverted microscope (IX71, Olympus) equipped with a 100×/1.45-NA oil-immersion objective. Cells were illuminated with high-power light-emitting diodes with wavelength of 405 nm (M405LP1, Thorlabs), 450 nm (M450LP1, Thorlabs) and/or 470 nm (M470 L3, Thorlabs). 405-nm and 470-nm illumination light were filtered at the exit of LEDs with band-pass filters centered at 405 nm (FBH405–10, Thorlabs) and 475 nm (FF01–475/28, Semrock), respectively. 450 nm illumination was filtered with a band-pass filters centered at 434 nm to allow the 440-nm component passing through. The fluorescence was collected through a 495-nm dichroic mirror (FF495-Di03, Semrock) and a 535/50 emission filter (FF01–535/50, Semrock). Illumination intensity was 5 mW/mm2 in all cases for 470-nm light and and 0.1~ 2.5 mW/mm2 for 405-nm light at the specimen plane. In all experiments, images were acquired continuously unless described explicitly, at 200 Hz with a sCMOS camera (Zyla, Andor). The camera and the microscope were connected with a 0.35×-magnification adapter (Olympus). Synchronization of electrophysiology and imaging was implemented with a DAQ board (PCI-6229, National Instruments) interfaced with Igor Pro (Wavemetrics) and Micro-Manager [20 (link)].
4
Plasmon-Enhanced Protein Imaging Setup
5
Holographic Imaging Setup for Fiber Bundles
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A schematic of the optical set-up is shown in Fig. 1. A 450 nm central wavelength, 15 nm bandwidth LED (Thorlabs M450LP1) was coupled into a 50 µm core, 0.22 NA multimode fiber which delivered light to the sample. The tip of the illumination fiber was approximately 15 mm from the tip of the fiber bundle (Fujikura FIGH-30-650S), and the sample was usually placed within 2 mm of the tip of the bundle. The bundle had an active imaging diameter of 600 µm, and contained approximately 30,000 cores arranged in a quasi-hexagonal pattern. The intensity of the inline hologram formed on the fiber bundle was transmitted in pixelated form and imaged onto a monochrome CMOS camera (Thorlabs DCC1545M) via a 20X objective and a 100 mm focal length tube lens. The magnification between the bundle and the camera was 11.8. With a camera pixel size of 5.2 µm, each pixel was projected to a size of approximately 0.44 µm at the bundle, and so the typical 3 µm inter-core spacing of the bundle was sampled by approximately 6 camera pixels. The image of the active area of the bundle was approximately 7.1 mm in diameter at the camera sensor plane. As the camera sensor was 6.66x5.32 mm this meant that the image of the circular bundle was slightly cropped.
6
Whole-Cell Patch-Clamp Electrophysiology
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Experiments were performed by EPC 800 patch clamp amplifier (HEKA Electronic GmbH, Pfalz, Germany). QDAS integrated and control devices were cleaned with 70 vol% ethanol solution and incubated for 3 days in DI water. The pulled patch pipettes of 4–6 MΩ were utilized to carry out the whole‐cell patch clamp experiment. The internal cellular medium was prepared by mixing 140 mm KCl, 2 mm MgCl2, 10 mm HEPES, 10 mm ethylene glycol‐bis (β‐aminoethyl ether)‐N,N,N′,N′‐tetraacetic acid, and 2 mm Mg‐ATP in water, and the pH was calibrated to 7.2–7.3 using 1 m KOH. The intracellular solution was filled into the patch pipettes to achieve whole‐cell patch. A digital camera integrated Olympus T2 upright microscope was used to patch and monitor the cells. The blue LED (M450LP1, Thorlabs Inc, NJ, USA) was used as the light source. LED system was driven by DC2200—High‐Power 1‐Channel LED Driver (Thorlabs Inc., NJ, USA).
7
Photoelectrochemical Characterization of Biointerfaces
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An Autolab Potentiostat Galvanostat PGSTAT system (Metrhom, Netherlands) was used for photoelectrochemical measurements, such as chronoamperometry, chronopotentiometry, cyclic voltammetry, and electrochemical impedance characterization. The three‐electrode system was used with a platinum rod as the counter electrode, Ag/AgCl as the reference electrode, and a connection to the ITO layer of the BI as the working electrode. 1 cm2 of the biointerface with the other two electrodes was immersed in an extracellular aCSF medium that mimics in vivo environment as an electrolyte solution at room temperature. The aCSF medium was prepared by mixing 10 mm of 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES), 10 mm glucose, 2 mm CaCl2, 140 mm of NaCl, 1 mm of MgCl2, and 3 mm of KCl. The pH of aCSF solution was adjusted to 7.4 by adding the required amount of NaOH solution. Then, a blue LED (M450LP1, Thorlabs) was used as the light source. The LED was driven by an LED Driver with Pulse Modulation (DC2200‐High‐Power 1‐Channel, Thorlabs) to produce a train of 20 ms light pulses. Then, the data was recorded and analyzed using the NOVA software. An optical power meter (Newport 843‐R) was utilized to measure the optical power densities of incident light on the biointerface.
8
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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