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Getting Started

This section will cover initial hardware and software setup. Please carefully unbox your equipment and install the PO5e card or UZ3 interface according to your System 3 Installation Guide. If you have a TDT WS4 or WS8 workstation, then a PO5e card will already be installed. Briefly - power down your computer* and place the PO5e card into an available PCIe slot in your computer. Next, install your TDT drivers and Synapse software from the USB Storage Drive that was provided with your shipment.

Note

* TDT drivers only function on Windows machines. Synapse will not run on Mac or Linux.

Important

This guide focuses on the RZ10(x) Fiber Photometry processor. If you are using the RZ5P or any other RZ processor, please refer to the Fiber Photometry User Guide for RZ5P Processor instead.

Below is a list of helpful online TDT resources with which users should be familiar before starting:

Synapse Training Videos
Narrated walk-throughs of the Synapse software. These are very helpful for beginner users first learning the Synapse environment.

Lightning Videos
Short, unnarrated videos that demonstrate specific actions in TDT software. These are referenced several times throughout this document, so look out for the blue icon

Knowledgebase
Contains documentation for all TDT hardware and software. This is a great first resource for troubleshooting

Tech Notes
Contain information about known hardware or software issues and asssociated solutions or workarounds

Support Help
TDT Tech Support offers phone and remote screen sharing support via GoToAssist to customers M - F, 8 AM - 5 PM Eastern Time. For remote screen sharing assistance, please email support@tdt.com to schedule an appointment.

Establishing RZ processor and PC communication

Once the PO5e card is seated and TDT drivers and software are installed, you are ready to connect the RZ processor (designated as RZ10(x) henceforth) and PC together. The orange fiber optic cables* will be used for PC-RZ communication (see Sys III manual for more details). Please connect the fiber optics to the correct ports on the RZ10(x) and PO5e card, as shown in the diagram below (red optical connector to 'Out' or Red-labeled ports on RZ and PC).

Note

* Your fiber optic cable may be a different length.

zBusMon with RZ10x Processor

Next, turn the RZ10(x) on. The display screen on the processor should illuminate with information about the unit's DSP cards (Run! u1 u2 u3...). To check whether there is communication between the RZ10(x) and the PC, open the zBusMon application (shown to the right). The RZ processor should appear with information about the driver version and number of DSP cards. Click Transfer Test to test communication.

If you get an error upon performing an initial transfer test, try performing a 'Reboot System!' first. If there is a consistent error in zBusMon, or you do not see your RZ appear, please contact TDT for assistance.

Launching Synapse

With your RZ10(x) on and connected, launch Synapse. The Rig Editor will appear, but it will be blank. Click Detect for Synapse to recognize your RZ10(x). For an RZ10x, a PC, RZ10x, three DSPs, and a PZ5 will show up in the tree. If you have a PZ5 preamplifier for recording electrical biopotential signals synchronized to your fiber photometry signal, this is where you would enable it by checking the PZ5(1) box. For an RZ10, a PC, RZ10 and one DSP will show up. If you have a Medusa4Z for up to four channels of biopotential data connected to the front legacy optics, you can add the device by right-clicking RZ10(x) → select Add RAn → click on the RA4PA A → change the Model → Medusa4Z. Finally, click Ok to exit the Rig Editor. The Rig Editor may be accessed later for modification through the Synapse Menu if your hardware changes.

The Rig Editor for modifications in Synapse Menu if your hardware
changes.

Your processor and any peripheral equipment declared in the Rig Editor will appear in the Processing Tree. For basic fiber photometry recordings, the experimental setup is simple. With the RZ10(x) selected, find the Fiber Photometry gizmo. Drag and drop, or double-click, the gizmo onto the RZ10(x) to form a connection.

Drag and drop fiber photometry gizmo for the RZ10(x) from TDT\'s
Synapse Software

Note

You must be running TDT Drivers and Synapse Version 94 or later.

You can learn more about gizmos and experimental connections in the Synapse Manual.

Detecting your fiber photometry equipment

Connection diagram for a 3-color fiber photometry setup.
The RZ10x is configured with 6 LEDs, 3 Photosensors, and 1 Power Meter

A general connection scheme for a 3-color fiber photometry setup is shown in the above diagram. RZ10x deluxe models have six LED light driver outputs and four sensor inputs organized into two banks. RZ10 base models have a single bank of three LED outputs and two sensor inputs.

Automatically Detect Connected LEDs and Sensors

The above RZ10x is configured with six Lux LEDs (405 nm, 465 nm, 560 nm), three Lux Photosensors, and one Lux Power Meter. For the 3-color setup, the Lux LEDs output light through a series of filters and dichroic mirrors ('fluorescent ports') that send excitation light to the subject and receive fluorescence back. The fluorescence signals are then sent to two Lux Photosensors on the RZ10x sensor inputs.

The RZ10(x) can also be configured with M8 output connectors to drive external LEDs, or BNC inputs to receive external photosensor signals. These can be interchanged by the user.

Important

In Synapse, on the Lux tab of the RZ10(x) gizmo, press the 'Detect Hardware' button. Synapse will automatically fill in the Driver (Drv-*) and Sensor (Sen-*) boxes based on the detected hardware. Possible options for the detected hardware include LED_{x}, M8 connector, PS1 photosensor, PM1 power meter, or BNC.

Performing a 'Detect Hardware' will automatically inform connected Fiber Photometry gizmos of the RZ10(x) configuration. See the Fiber Photometry Gizmo section for more details.

LED_{x} - This is a Lux LED of a specified wavelength x. Common wavelengths used in fiber photometry include 405 nm (autofluorescence detection, isosbestic control), 465 nm (GCaMP, dLight), 560 nm (TDtomato, mCherry, RCaMP). Please see the Lux LED webpage for a list of all available wavelengths.

M8 - This is an M8 connector that is commonly used for external LEDs. Standalone LEDs from Thor Labs and Doric both use M8 connectors for power.

PS1 - This is the Lux photosensor.

PM1 - This is the Lux power meter.

BNC - This is a BNC (coaxial) connector that can be used to drive an external LED driver or receive the output of an external photoreceiver. This connector enables the 'DAC Out' or 'ADC In' checkbox, depending on if the BNC is for the Driver or Sensor hardware slots. Enable this checkbox only if you are using the BNC connector outside of the Fiber Photometry gizmo. It will be available on the 'DAC' and 'ADC' tabs, respectively.

Fluorescent Ports - these are the series of filters and dichroic mirrors that send excitation light to the subject and receive fluorescence back. Many labs will use Doric Mini Cubes as their light filters instead of creating their own optical benchtop, but both options are feasible. These need to be configured specifically for the wavelengths of light sources and fluorescent signals that are expected. Be sure to route the appropriate light wavelengths to the correct bandpass filter ports.

For example: with a 465 nm GCaMP + 405 nm isosbestic setup that uses a four-port Doric Minicube, the 465 light will route to E1, the 405 light to AE, the subject will be connected to Sample, and the output to the photosensor will be the F1 port.

Fiber optic patch cords - TDT sells a fiber optic patch cable kit with our recommended cables. This includes: a 200 µm core diameter cables for the LED to fluorescent port/ Minicube connection; a 600 µm core diameter cable for the fluorescent port/ Minicube to PS1 connection; a 400 µm core diameter cable to serve as the Subject cable* when connected to the Lux PM1 power meter. All cables should have a black jacket to prevent ambient light interference. TDT also recommends that customers order low auto-fluorescent specific subject cables from either Doric or Thorlabs.

Note

* For accurate power measurements for your setup, the core diameter of the PM1 cable should match the core diameter of the Subject cable that you are using in your experiments (typically either 200 µm or 400 µm).

For customers who want to use larger core diameter cables, such as 400 µm, but need to drive power levels low (less than 40 µW), TDT sells an 85% attenuation coupler to reduce the amount of light going to the subject. The attenuation coupler connects as follows: LED Patch Cable Attenuation Coupler Patch Cable Fluorescent Port/ Minicube.

For setups with external LED drivers (especially Doric) and a TDT RZ5P, it was common to use patch cords with attenuation filters (1%, 5%, or 10%) to reduce the power output of the excitation light sources before light reaches the fluorescent ports. This is because Fiber photometry is a low light power application, and it was often difficult to drive the LEDs with low enough currents to reach target power levels. The RZ10(x) has superior output signal quality and can adjust the max current output range to allow for very low current outputs, so using attenuating patch cables is not necessary. Also, never connect an attenuating fiber to the photoreceiver; this will severely diminish fluorescent output.

If Using a 3rd Party Photosensor - this would be connected to a LUX BNC connector in place of the PS1. For Doric or Newport photoreceivers, the gain should always be set to DC Low. This provides the widest bandwidth of light detection and detects signal clipping easier. Here is a link to the photoreceiver frequency response plots. If your photoreceiver has a 1x, 10x, 100x option, typically 10x will provide the clearest output response.

Adding a USB Camera

Configuration of low frame rate (20 fps or less) subject monitoring via USB cameras is simple in Synapse. Cameras can be added in the Rig Editor. Please follow this Lightning Video for specific instructions.