Fiber Photometry for RZ10 Processor
Common Use Cases
Real-time control and acquisition of demodulated locked-in amplification signal from any combination of up to 3 light drivers and 2 photosensors on a Lux bank. Can also monitor light power from PM1 from any Lux bank. This is the primary gizmo used in fiber photometry setups using RZ10 or RZ10x processors. Record up to 6 demodulated signals with raw photosensor output and dF/F calculations, too.
Gizmo Help Slides
The Fiber Photometry gizmo includes designtime and runtime control for the RZ10 LuxIO processor built-in LED light drivers. It uses lock-in amplification to measure the real-time power of the resulting fluorescence response at the LED driving frequencies.
It provides an estimate of the LED power output and can also measure power at the subject end. It has a built-in timer for timed stimulus control and automatic photo-bleaching capabilities.
Each Fiber Photometry gizmo targets a single LUX bank of components on the RZ10, controlling up to three LED drivers and reading two photosensors. PM1 power meters are handled uniquely; they can be accessed from any LUX bank.
The Runtime Interface
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 (405A, 465A) at the frequencies of the 405 and 465 driver signals on the sensor A input. The Fiber Photometry gizmo also stores and displays broadband raw input signals and driver parameters, depending on selections made at designtime. Fi1r is the raw sensor input in this example.
Note that the Power Meter and Fiber Bleaching modes are only available in Preview mode. Power Meter mode is for system verification, and Fiber Bleaching is a special feature that doesn't involve data collection, so these modes can't be accidentally enabled during an experiment (Record mode).
|Runtime Interface for Power Meter Mode|
Note that the Power Meter mode is only available when a LUX PM1 is detected in the RZ10. This mode is for system verification and is only available in Preview mode, so it can't be accidentally enabled during an experiment (Record mode).
|Runtime Interface for Fiber Bleaching Mode|
The Fiber Bleaching mode is a special feature for driving high light power through the cables for an extended period of time to reduce the cable's autofluorescence and increase signal to noise before running an experiment. This mode doesn't involve data collection so it is only available in Preview mode, so these modes can't be accidentally enabled during an experiment (Record mode).
Check with your fiber optic cable manufacturer for recommendations on fiber bleaching current, duration, and how often this should be done.
Fiber Photometry Configuration Options
Lock Freqs at Runtime
Uncheck this for debugging or system setup, otherwise leave it unchecked to prevent accidental frequency changes during recording.
Automatically sets the Offset to 10% of the Level (rounded up) at runtime, with a minimum of 5 mA and maximum of 20 mA. This can reduce signal distortion at higher Level settings.
Launch Power Est
This option will display an estimate of the LED power output (in uW) at runtime through the selected fiber core diameter connected to the LED, assuming the cable Numerical Aperture is 0.48. Use this setting with a PM1 power meter to get an estimation of overall light transmission through the entire optical chain.
Note: PM1 power meters do not appear in this tab but are available in the runtime interface in Preview mode.
Set the storage sampling rate for the demodulated and calculated signals configured on this tab.
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.
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.
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.
Lux Options Tab
|Lux Options Tab|
Timing control options are used to cycle the LEDs On and Off for set durations and repeats during runtime. This feature is for running long (greater than 1 hour) experiments where photobleaching becomes a concern.
This option group is only available if a PM1 is detected in the RZ10(x).
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
Check this option if you want to automatically turn on the LED drivers when switching to Preview or Record mode.
Store Driver Signals
Stores the signals used to drive the LEDs at the RZ10 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 RZ10 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.