Donka Rachel M, Loh Maxine K, Konanur Vaibhav R, Roitman Mitchell F, Roitman Jamie D
Department of Psychology, University of Illinois Chicago, Chicago, Illinois.
Graduate Program in Neuroscience, University of Illinois Chicago, Chicago, Illinois.
Curr Protoc. 2025 Jul;5(7):e70161. doi: 10.1002/cpz1.70161.
Fiber photometry is a rapidly growing technique to record real-time neural signaling in awake, behaving subjects. However, the processing and analysis of photometry data streams can be complicated, and there is wide divergence in methods across the field. Although several open-source signal processing tools exist, platforms can be inflexible in accommodating experimental designs, lack consistency in signal peak detection, and difficult to use for novice users. In addition, more recent sensors produce fluorescence that may be detected on both signal and control streams, confounding signal processing. To remedy these challenges, we developed PASTa (Photometry Analysis and Signal Processing Toolbox), an open-source MATLAB-based toolbox and protocol for the processing and analysis of fiber photometry data. PASTa includes a full analysis pipeline from data preparation through signal processing and transient event detection. Default parameters were selected to provide users with a conservative starting place, with optional inputs to include other commonly used methods and techniques in the field. Additionally, the transient detection protocol adopts options for determining peak detection thresholds and pre-peak baselines to allow more reliable detection of events and characterization of transient kinetics. Although operating through MATLAB, the code is annotated to be readable, accessible for new users, and adaptable. Ultimately, the PASTa protocol introduces a customizable, user-friendly platform to process fiber photometry signals and detect and quantify transient events. Novel methods for the analysis of signal processing will continue to be integrated into the toolbox, seeking to provide fiber photometry users with access to a comprehensive analysis suite that can accommodate the wide diversity of sensors and experimental paradigms used in photometry. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Software installation Basic Protocol 2: Data preparation Basic Protocol 3: Signal processing Basic Protocol 4: Transient detection and analysis Support Protocol: Function default customization.
光纤光度法是一种在清醒、行为活动的受试者中记录实时神经信号的快速发展的技术。然而，光度学数据流的处理和分析可能很复杂，并且该领域的方法存在很大差异。尽管存在几种开源信号处理工具，但这些平台在适应实验设计方面可能不够灵活，在信号峰值检测方面缺乏一致性，并且对于新手用户来说难以使用。此外，更新的传感器产生的荧光可能在信号流和对照流中都能被检测到，这会干扰信号处理。为了应对这些挑战，我们开发了PASTa（光度分析和信号处理工具箱），这是一个基于MATLAB的开源工具箱和协议，用于处理和分析光纤光度数据。PASTa包括从数据准备到信号处理和瞬态事件检测的完整分析流程。选择默认参数为用户提供一个保守的起点，并提供可选输入以纳入该领域其他常用的方法和技术。此外，瞬态检测协议采用了确定峰值检测阈值和峰前基线的选项，以便更可靠地检测事件并表征瞬态动力学。尽管通过MATLAB运行，但代码添加了注释，易于阅读，新用户可以访问并进行修改。最终，PASTa协议引入了一个可定制、用户友好的平台来处理光纤光度信号，并检测和量化瞬态事件。信号处理分析的新方法将继续集成到该工具箱中，旨在为光纤光度用户提供一个全面的分析套件，以适应光度学中使用的各种传感器和实验范式。© 2025作者。由Wiley Periodicals LLC出版的《当前协议》。基本方案1：软件安装 基本方案2：数据准备 基本方案3：信号处理 基本方案4：瞬态检测与分析 支持方案：函数默认定制
