An objective-type TIRF microscope was set-up, using a Nikon TiE-inverted microscope equipped with a perfect focusing unit, bright-field illumination and a × 100 1.49 numerical aperture PlanApo oil immersion objective (Nikon). A 488-nm solid state laser (coherent) was used for GFP excitation. The GFP signal was recorded by an Andor iXon 512 × 512 electron-multiplied charge-coupled device (EM-CCD) camera. Extra magnification (× 1.5) was used to obtain a pixel size of 106 nm. Excitation laser beams were controlled by Uniblitz shutters. Because the CCD image is saturated under intense laser illumination, shutter timing was synchronized with the camera acquisition by a DAQ card (NI, USB-6221). In a continuous acquisition mode of the camera, frames that were exposed to a high-power PhotoGate beam was not used for data analysis. In a time-lapse data acquisition mode, the gate beam was turned on during the times when the camera was not acquiring data.
Usb 6221
Manufactured by National Instruments
Sourced in United States
The USB-6221 is a multifunction data acquisition (DAQ) device from National Instruments. It provides 16-bit analog input channels, 2 analog output channels, and 24 digital I/O lines. The device connects to a computer using a USB interface.
Automatically generated - may contain errors
Lab products found in correlation
Ti e inverted microscope, by Nikon (1 mentions)
Plan apo oil immersion objective, by Nikon (1 mentions)
Ni daqmx, by National Instruments (1 mentions)
Bagnoli 8 emg system, by Delsys (1 mentions)
2.1 single differential configuration, by Delsys (1 mentions)
Labview, by National Instruments (1 mentions)
Pcie 6361, by National Instruments (1 mentions)
Usb 6229, by National Instruments (1 mentions)
Usb 6001, by National Instruments (1 mentions)
Sr570, by Stanford Research Systems (1 mentions)
5 protocols using usb 6221
1
Single-molecule TIRF Microscope Setup
2
Multichannel Facial EMG Recording
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Four EMG channels were recorded, from Mentalis, left Risorius, right Risorius, and left Temporalis muscles. The skin was first cleaned using alcohol wipes, and EMG sensors (Delsys 2.1 single differential configuration, Delsys Inc., Boston, USA) were placed over the muscles using double-sided adhesive tape. The reference electrode was placed on right clavicle. The raw EMG signals were filtered (20–450 Hz band pass filter) and amplified 10,000 times by a Delsys Bagnoli-8 EMG System (Delsys Inc., Boston, USA). The EMG signals were digitized at a sampling rate of 1,000 Hz by a 16-bit analog-to-digital converter (USB-6221, National Instruments Inc., Austin, USA), recorded on a computer (Windows 7, Intel Celeron 1007 U at 1.5 GHz with 4 GB RAM) via USB, and accessed using a National Instruments programming interface (NI-DAQmx) for C++.
3
Head-fixed Behavioral Training in Mice
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After recovery from the head-plate implantation, the mice were water-deprived in their home cages. They received about 1 mL water per session every day, and were sometimes given additional water to maintain their body weight at 80–85% of their initial weight throughout the experiments. The mice were usually trained for five consecutive days per week, and were given a 1.2–1.4 g agar block (Oriental Yeast Co., Ltd., Itabashi, Tokyo, Japan) on days without training. The behavioral apparatus (sound attenuation chamber, head-fixing frame, body holder, sound presentation system, water-supply system, and integrated lever device) was manufactured by O’hara & Co., Ltd. (Nakano, Tokyo, Japan). The lever position was monitored by a magnetic sensor and was continuously recorded at an acquisition rate of 1000 Hz by a NI-DAQ (USB-6001, USB-6221, USB-6229, or PCIe-6361; National Instruments, Austin, TX, United States). The sound control and water delivery were controlled using a program written in LabVIEW (National Instruments).
4
Optogenetic Seizure Modulation Protocol
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Light was delivered 6–7 days following viral injection. For light delivery, the optical fiber implant was connected to a 473 nm diode-pumped solid-state laser (Shanghai Laser & Optics Century). Mice were allowed 2 h to recover from implant surgery before light delivery. For kainate experiments, closed-loop seizure detection and light delivery were carried out as previously described22 (link). Briefly, LFP recording electrodes (PlasticsOne) for kainate-injected animals were connected to an electrical commutator (PlasticsOne) routed to an amplifier (BrownLee 410, Automate Scientific), and in turn connected to a digitizer (USB-6221, National Instruments) and a computer running custom MATLAB recorder and seizure detection software. When a seizure was detected, the software enabled light delivery. Animals in all groups receiving light had one single session of 10 mW 473 nm light delivered in 2 s pulses every 6 s (33% duty cycle), for a total of 10 min. Animals were euthanized and perfused 18–24 h after the end of light administration.
5
Coaxial Relay Switch for I(V) and Thermovoltage
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A coaxial relay switch was used to switch between I(V) and thermovoltage mode. In I(V) mode voltage is sourced by a data acquisition (DAQ) card (National Instruments, USB 6221) (linear ramps: 0-max–min-0). Current was preamplified by an I–V converter (Stanford Research Systems, SR570) and measured via DAQ card. Acquisition times of I(V) curves were 0.5 s. These short measurement times were chosen to make sure that successive I(V) curves can describe the same structures (I(V) measurement time < lifetime of junctions). In thermovoltage mode the relay was opened, creating open-contact conditions. Thermovoltages were amplified via a pre-amplifier (FEMTO, DLPVA) and digitized via DAQ card. Offset currents caused by the pre-amplifier were compensated using a source measure unit (Agilent, E5287A) in current source mode (offset current constant throughout measurement). This increased the trusted conductance range from to (offset current decreased from 10 pA to 1 pA). A circuit diagram and more detailed description can be found in SI.
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