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Fibre Photometry

Plug&play assembly service 

Data acquisition software

We assemble the fibre photometry systems using components from Doric Lenses and pair it with our custom acquisition software.

See below for free build instructions and download. 

 

Purchase list (last updated March 2023)

 

1Low autofluorescent options are preferred. Silica/polymer is fine.

2NA of the fibre optics [animal  cube  detector] must be kept consistent (600mm core not possible with 0.57NA)

3Newport 2151 not required if built-in detector is purchased

4The Newport 2151 is an excellent photosensor for fibre photometry. It is used across most of our photometry rigs and we see no reason to purchase any other detector.

5We generally have very little noise issues with the standard rotary joints. They offer better light throughput, replaceable optics, and less likelihood for damage over the pigtailed models. We bleach-off the autofluorescence within our rotary joints prior to experiments.

6This is a consumable and is prone to damage over use

Computer specifications: 

Windows 10 or 11.

Minimum requirements: SSD (at least 256gb), 8gb Ram, and i5 processor.

Additional storage is recommended. 

Photometry system hardware assembly instructions 

1. Our acquisition software controls LED modulation and power via the National Instruments (NI) board. The emitted fluorescence is detected by the Newport 2151 and subsequently acquired via the NI board. Connections between the LED driver and photodetector with the National Instruments are as follows

  • Doric Driver LED1 Input to NI Analog Output #12 - See A in image below

  • Doric Driver LED2 Input to NI Analog Output #13 - See B in image below

  • Common ground to Analog Output #14 

  • Optional Analog output #4 for TTL trigger 

  • Newport 2151 BNC Out to NI Analog Input #28 - See C in image below

  • Ground to Analog Input #27

2. USB connections to computer. 

The Doric LED driver and the NI board both require USB connections to the computer. Power adjustments and low power modes are set via USB command codes to the LED driver. The acquisition software also checks that the LED driver is present and is connected to the correct LEDs. NI board connection is required for LED modulation and data acquisition. 

Argotech can provide assembly services to create an all-in-one photometry box. With single power and usb ports and optical output for a plug and play system.
 

 

3. LEDs and Internal optics

Connect the excitation (465nm) LED to LED#1 and isosbestic control (405 or 410nm) LED to LED#2. The LEDs should then be connected to the mini cube. The emitted green fluorescence from the mini cube is connected by optocal cable to the Newport detector. See purchase list for optical cable specifications. 

Data acquisition software installation 

1. Download the free software and follow the installation instructions. If this is the first installation, please continue the steps to install the National Instruments 1550f Runtime. The Runtime should only be installed the one time upon first install. 

 

Acquisition softwares v1.08 and v1.09 are available below. 

If your Doric LED driver has the newest firmware installed, please install v1.09 as it requires new sets of USB command codes. 

If you are not sure which version you need, connect your computer to the LED driver and open Device Manager. If your driver is recognised as “LightSource Driver”, you have the latest firmware and will need v1.09. If the driver is recognised as "Optogenetics System", you will need v1.08. Please note that v1.09 is not compatible with previous generations of LED driver and vice versa. 

2. Open the fibre photometry software. Before acquisition, click "Settings" on bottom left corner and set LED power and directory for saved data. The software automatically saves raw data during acquisition. For duty cycle mode, set your desired on/off durations for the LEDs and set "enable". 

3. To begin data acquisition, click "Start" on bottom right corner. The 3 windows will display real time demodulated emitted data from the excitation, the isosbestic control LEDs, and subtraction of the two. Note that the subtraction window does not use the scaled 405nm signal and may not be accurate. 

4. To finish, click "Stop" and export the demodulated data. Click, file/export. This will export 10Hz demodulated data from the two wavelengths into a single csv file. 

 

 

 

 

 

 

 

Blueprints of the fibre photometry system and daq:

 

 

 

 

 

 

 

 

 

 

 

Data analysis and DF/F calculations 

Coming soon: Custom python script (based from GuPPY) for DF/F and Z-score calculations will be uploaded shortly.  

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