LabVIEW-DAQ

LabVIEW VIs for whisker tracking, force sensor analysis, and other behavioral measurements to supplement animal arousal-state characterization during IOS imaging and 2PLSM.

This project is maintained by KL-Turner

LabVIEW-DAQ VIs and Hardware

LabVIEW VIs for whisker tracking and force sensor analysis to supplement animal behavioral characterization during two photon microscopy

https://kl-turner.github.io/LabVIEW-DAQ/

LabVIEW VIs for behavioral characterization for both Two Photon and Intrinsic Optical Signal imaging set-ups. This README will break down the different components and give a brief run-down of the respective parameters. The documentation, user manuals, diagrams, and spec sheets linked in this repository are freely available online or upon request. The company names and product names outlined in this README are trademarks, registered trademarks, or copyrights of the respective companies.

This repository has two main folders that hold the project files and associated subVIs for the Two Photon and IOS components. To open the Front panel, naviate to either fold and either open the labview project file(.lvproj) or the MasterAcquisionFile_[2P/IOS] file. I recommend opening the MasterAcquitionFile directly. Do not be overwhelmed by the block diagrams - Ideally, much of the Master VI’s content would be subdivided into smaller more dedicated VIs, however, due to the re-iterative nature of these projects and how the legacy versions of these codes were originally written, the majority of the project’s contents lies within the Master VI even though this is against good (graphical) coding practices.

Two-Photon Excitation Microscopy

MasterAcquisionFile_IOS Front Panel

This VI runs in synchony with MScan, starting from a TTL trigger sent by MScan’s scanning initiation. When run, the VI will initiate all of the hardware paths to reduce the lag time between MScan and the VI starting. It will then stand in a while loop until the designated pin receives the trigger, ending the loop and entering the DAQ loops. A Notes section for experimental parameters and any analog signals will be saved to a .tdms file with a .tdms_index file giving the parameter indeces. Any camera files are saved independently with their own respective (typically .bin) extensions. The force sensor signal is duplicated to both MScan and LabVIEW so that any remaining delay between the two can be corrected in-post.

Numerous indicator lights signal the current status of the program. Once DAQ starts, the webcam (large rectangle) and whisker camera (narrow rectange) will give real-time monitoring of the camera feeds. Numerous monitors of the analog samples, whisker camera sampling rate, and web cam sampling rate allow the user to troubleshoot any issues. A dropped-frames meter will monitor the dropped whisker camera frames. Note that the webcam is just for general purposes, and is actually not being triggered. The webcam’s settings, depending on the model, can be modulated in NI MAX. While many webcams are IR sensitive, nearly all have a filter that can be removed. This is important because any significant amounts of light will pop the PMTs/ruin your image. We use a webcam as opposed to a more expensive camera because the movies are smaller in size and are only used for strange events and real-time monitoring, not in any significant data analysis. There are numerous controls for folder paths and hardware/camera IDs. For any questions regarding these, consult your respective National Instruments Device documentation and NI MAX settings.

Equipment

Hardware, Manufacture Documentation Purpose
Spectra-Physics Mai Tai Ti:Sapphire laser https://www.spectra-physics.com/products/ultrafast-lasers/mai-tai Two-photon laser
National Instruments PCIe-6353 http://www.ni.com/en-us/support/model.pcie-6353.html Integrates various components with MScan
National Instruments PCIe-6321 http://www.ni.com/en-us/support/model.pcie-6321.html Integrates various components with MScan
National Instruments BNC-2110 (x4) http://sine.ni.com/nips/cds/view/p/lang/en/nid/1865 Integrates various components with MScan
National Instruments USB-6211 http://www.ni.com/en-us/support/model.usb-6211.html DAQ for webcam, force sensor, whisker cam
Sutter Instruments PS-2LV https://www.sutter.com/MICROSCOPES/pmt_frame.html PMT power supply
Sutter Instruments MDR-6 See new Resonant version MDR-R https://www.sutter.com/MICROSCOPES/resscan.html Scan drive controller
Sutter Intruments MPC-200 https://www.sutter.com/MICROMANIPULATION/mpc200.html Objective micromanipulator
Conoptics Model 350-80LA https://www.conoptics.com/modulation-systems-mpm/ Electro-Optic modulator, control laser intensity
Conoptics Model 320RM https://www.conoptics.com/modulation-systems-mpm/ Pockel cell controller
Uniblitz VCM-D1 https://www.uniblitz.com/products/vcm-d1-shutter-driver/ Shutter driver and controller
Femto DHPCA-100 (x2) https://www.femto.de/en/products/current-amplifiers/variable-gain-up-to-200-mhz-dhpca.html Variable gain high-speed current amplifier for PMTs
Nikon CFI75 LWD 16X W Objective https://www.nikon.com/products/microscope-solutions/lineup/objectives/cfi75_water_dipping_series/index.htm Water immersion objective
Thorlabs LEDs M780L3 (x2) https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=2692&pn=M780L3#4426 2 Infrared for whisker/webcam
Thorlabs LEDD1B T-Cube LED Driver (x5) https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=2616 Power and illumination control for Thorlabs LEDs
Newport KPX124 Plano-Convex Lens (x1) https://www.newport.com/p/KPX124 General lens for Thorlabs LEDs
Basler ace acA640-120gm (camera) https://www.baslerweb.com/en/products/cameras/area-scan-cameras/ace/aca640-120gm/ Whisker tracking camera
Edmund Optics 35mm C Series VIS-NIR FFL Lens https://www.edmundoptics.com/p/35mm-c-series-vis-nir-fixed-focal-length-lens/22384/ Lens for whisker camera
Tekscan FlexiForce quickstart board https://www.tekscan.com/products-solutions/electronics/flexiforce-quickstart-board Force sensor for movement detection
ELP 2.8mm wide angle IR LED Infrared USB camera https://www.amazon.com/gp/product/B01E8OX14K/ref=ppx_yo_dt_b_asin_title_o01_s01?ie=UTF8&psc=1 General purpose webcam
RW Automation Solenoid Control Board SC5 http://www.rwautomation.com/ Control board for whisker, auditory stimulation
STC Solenoid Valve 2V035-1/4 (x3) https://www.stcvalve.com/Solenoid-Valve/2V025-035.htm Solenoid valve that directs 10 psi of air
World Precision Instruments DAM80 (X2) https://www.wpiinc.com/sys-dam80-extracellular-amplifier-with-active-probe Diff Amp for 1 neural electrode, 1 EMG
Stanford Research Systems Model SR560 https://www.thinksrs.com/products/sr560.html Preamplifier for force sensor
TENMA Dual Power supply 72-8690A (x2) https://www.newark.com/tenma/72-8690a/power-supply-2ch-32v-3a-adj-fixed/dp/47X1649 Power for Basler camera, solenoids, force sensor

See https://www.edmundoptics.com/search/?criteria=mirror%20coating for mirror options to direct laser path.

MasterAcquisionFile_2P Block Diagram
Two-photon set-up (portable) on cart
Full Two-Photon set-up

Intrinsic Optical Signal (IOS) Reflectance Imaging

MasterAcquisionFile_IOS Front Panel

This VI runs independently and acquires all data itself. It shares all the same monitoring and control attributes as the Two Photon VI described above, as well as several more. In addition to the webcam and whisker camera feeds, this VI also shows the pupil diameter and IOS window in real-time.

Equipment

Hardware, Manufacture Documentation Purpose
National Instruments PCI-6259 http://sine.ni.com/nips/cds/view/p/lang/en/nid/14128 Device interface for DAQ and counter outputs
National Instruments BNC-2090A http://sine.ni.com/nips/cds/view/p/lang/en/nid/203462 BNC interface for PCI-6259
National Instruments USB-6343 http://www.ni.com/en-us/support/model.usb-6343.html Additional counter output
National Instruments PCI-1428 http://www.ni.com/en-us/support/model.pci-1428.html Framegrabber for DALSA 1M60 camera
Teledyne DALSA 1M60 (camera) https://www.teledynedalsa.com/en/products/imaging/cameras/ IOS reflectance camera
Edmund Optics VZM 300i Zoom Imaging Lens https://www.edmundoptics.com/p/vzmtrade-300i-zoom-imaging-lens/10634/ Lens for IOS camera
Edmund Optics M22.5x0.5 Green Filter https://www.edmundoptics.com/p/mounted-m225-x-05-threaded-green-filter/6901/ Bandpass filter for IOS camera
Thorlabs LEDs M530L3 (x4), M780L3 (x1) https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=2692&pn=M780L3#4426 4 green IOS LEDs, 1 Infrared for pupil tracking
Thorlabs FB530-10 Bandpass filter (x4) https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=2616 Bandpass Filter for IOS Thorlabs LEDs
Thorlabs LEDD1B T-Cube LED Driver (x5) https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=2616 Power and illumination control for Thorlabs LEDs
Newport KPX076 Plano-Convex Lens (x5) https://www.newport.com/p/KPX076 Columnating lens for Thorlabs LEDs
Basler ace acA640-120gm (camera) https://www.baslerweb.com/en/products/cameras/area-scan-cameras/ace/aca640-120gm/ Whisker tracking camera
Edmund Optics 18mm DG Series FFL Lens https://www.edmundoptics.com/p/18mm-dg-series-fixed-focal-length-lens/11591/ Lens for whisker camera
Edmund Optics 50x50mm 660nm LED backlight https://www.edmundoptics.com/p/50-x-50mm-660nm-led-backlight/21332/ Illuminates whiskers for whisker camera
Edmund Optics 75 mm DG Series FFL Lens https://www.edmundoptics.com/p/75mm-dg-series-fixed-focal-length-lens/11371/ Lens for pupil camera
Tekscan FlexiForce quickstart board https://www.tekscan.com/products-solutions/electronics/flexiforce-quickstart-board Force sensor for movement detection
Microsoft Lifecam Cinema https://www.microsoft.com/accessories/en-us/products/webcams/lifecam-cinema/h5d-00013 General purpose webcam
RW Automation Solenoid Control Board SC5 http://www.rwautomation.com/ Control board for whisker, auditory stimulation
STC Solenoid Valve 2V035-1/4 (x3) https://www.stcvalve.com/Solenoid-Valve/2V025-035.htm Solenoid valve that directs 10 psi of air
World Precision Instruments DAM80 (x3) https://www.wpiinc.com/sys-dam80-extracellular-amplifier-with-active-probe Diff Amp for 2 neural electrodes, 1 EMG
Brownlee Precision Model 440 Amplifier http://www.brownleeprecision.com/products/amplifier-model-440/ Amplification of force sensor, filter MUA for audio
Tektronix TDS 2014C Oscilloscope (x2) https://www.tek.com/oscilloscope/tds2000-digital-storage-oscilloscope Monitor signals in real time
TENMA Dual Power supply, 280W (x2) http://www.tenma.com/ Power for Basler cameras, solenoids, force sensor

Both imaging systems are built with numerous Thorlabs components, in particular https://www.thorlabs.com/navigation.cfm?guide_id=50 on vibration-isolation optical tables.

MasterAcquisionFile_IOS Block Diagram
IOS Setup

Acknowledgements

Feel free to contact Kevin Turner (klt8@psu.edu) with any questions regarding the hardware or VIs.