Fiber Photometry for non-RZ10x Processors

Common Use Cases

Real-time control and acquisition of demodulated locked-in amplification signal from any combination of up to 4 light drivers and 2 photosensors. This is the primary gizmo used in fiber photometry setups using processors other than RZ10x. Record up to 8 demodulated signals with raw photosensor output too.

Data Stored
Stream	Demodulated response signals
Stream (optional)	Broadband raw signals
Scaler (optional)	Driver parameters
Outputs
Driver output voltage (optional)	Up to 4 single-channel floats
Calculated signals (optional)	Up to 4 single-channel floats

Gizmo Help Slides

Quick Help Slide 1Quick Help Slide 2

Reference

The Fiber Photometry gizmo, when used with processors that are not the RZ10x, includes designtime and runtime control of up to four voltage output signals to drive external light drivers. It uses lock-in amplification to measure the real- time power of the resulting fluorescence response at the light driving frequencies. It controls up to four light driver signals and reads two photosensors via the RZ analog inputs.

The Runtime Interface

Runtime Plot

A plot is added to the runtime window for visualization.

Flow Plot Showing Demodulated Responses

The subplots shown in a runtime plot represent data storage you chose in the designtime options. In the example above, the streamed data shows the resulting power output (such as Dv1A) at the frequency of interest when comparing the selected driver (such as Drv1) to the selected sensor input (such as sensor A). Simultaneous neural recordings from a different gizmo are integrated in the plot for a quick visual comparison. The Fiber Photometry gizmo also stores and displays broadband raw input signals and driver parameters, depending on selections made at designtime.

Runtime Controls

Runtime Interface

The runtime window includes:

Options	Description
Clipping Indicator(s)	Two indicators, one for each sensor, flash when the user-defined clipping threshold is crossed. The clipping indicator will also light up if the input voltage is below 10 uV to indicate a bad connection.
Lowpass Filter	A knob and value entry box that controls the lowpass filter that determines the bandwidth around each demodulation frequency used for the power calculation
Dv{x} Options
Light On	A button enables the light driver and an indicator is lit green when the light is on
Parameters	Knobs and value entry boxes allow runtime control of light driver Frequency, Level, and DC Offset parameter values
Results	A result for each sensor is dynamically displayed as a single value in millivolts

Fiber Photometry Configuration Options

See The Options Area and Templates for more information on the gizmo name, source, global options, and displaying the block diagram.

Driver(s) Tab

Drv-{n} Options	Description
Name	Three characters that serve as the first three characters in the store name.
Output	Specify the DAC number (channel number in parenthesis), or send this as a gizmo output.
Cal Factor	Scale factor used to convert desired milliamps into voltage.
Defaults Options
Frequency	Modulation frequency of the light source sine wave
Level	Peak-to-peak light source sine wave amplitude
Offset	DC shift added to the output sine wave. The offset is adjusted to reach the linear range of the physical light driver output and minimize signal distortion. More on that later.

Sensor(s) Tab

Sensor Option	Description
Name	Give the sensor a name as it will appear in the runtime interface. The first letter of the sensor name is used as the last letter of the streaming data store name.
Source	The sensor input can be an analog input from the front panel of the hardware or a floating point signal output of another hardware source or gizmo.
Calibration Factor	Scales the sensor data
Clip Threshold	Raw A/D sensor input voltage value to light runtime indicator (no calibration factor applied)
Sensor B check box	Enables a second sensor input
Demodulator Option
Filter Order	Higher order filters tighten the band around the response frequency
Default Lowpass Frequency	Determines the band around the frequency of interest to do the demodulation calculation. This can be modified at runtime.
Enabled Higher Lowpass Range	Check this box to raise the lowpass frequency limit to 100 Hz for TEMPO (voltage sensor photometry) where sensors have a wider bandwidth.

Demodulator(s) Tab

Storage Rate

Set the storage sampling rate for the demodulated and calculated signals configured on this tab.

Demodulator Options

Use the matrix of check boxes to select the combinations of sensors and drivers that will be used for demodulation. All available sensor signals can be demodulated against all light driver signals if desired.

Calculated Outputs

Perform up to four real-time calculations on the demodulated data streams. Choose a Source demodulated signal, optionally subtract another demodulated signal in the Difference with... column, and optionally perform a delta F over F calculation on it (dF/F checkbox) to compare the strength of the signal relative to the baseline.

Saving

The calculated signal is available as a gizmo output so other gizmos can attach to it and do further processing in real-time. The calculated signal can also be optionally shown on the Flow Plot at runtime, and optionally be saved to disk as well.

dF/F Options - Default Window Duration

Sliding average window size used as the baseline for the delta F over F computation.

Misc Tab

Required Sample Rate

Tells the RZ what minimum sample rate this gizmo requires. Typically 6K is enough. Only increase this if the driver frequency needs to go beyond 1-2 kHz for your experiment, which is rarely done.

Drivers On at Runtime

When selected, output driving signals are "on" when recording begins. Otherwise, light drivers must be turned on manually in the runtime interface.

Store Driver Signals

Stores the signals used to drive the LEDs at the RZ system rate. The store name is the first two letters of the gizmo name, followed by the last letter of gizmo name, followed by 'd' (default 'Fi1d'). One channel per light driver.

Store Driver Parameters

All light driver parameters are timestamped and stored to disk two seconds after a change has been made to any of the driver parameters in the runtime user interface.

Store Sensor Signals

Stores the raw sensor signals at the RZ system rate. The store name is the first two letters of the gizmo name, followed by the last letter of gizmo name, followed by 'r' (default 'Fi1r'). One channel per sensor.