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Research ArticleResearch Article: Methods/New Tools, Novel Tools and Methods

FiPhoPHA - A fiber photometry python package for post-hoc analysis

Vasilios Drakopoulos, Alex Reichenbach, Romana Stark, Claire J. Foldi, Philip Jean-Richard-dit-Bressel and Zane B. Andrews
eNeuro 29 July 2025, ENEURO.0221-25.2025; https://doi.org/10.1523/ENEURO.0221-25.2025
Vasilios Drakopoulos
1Monash Biomedicine Discovery Institute and Department of Physiology, Monash University, Clayton, Victoria, Australia.
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  • For correspondence: Vasilios.Drakopoulos{at}monash.edu Zane.Andrews{at}monash.edu
Alex Reichenbach
1Monash Biomedicine Discovery Institute and Department of Physiology, Monash University, Clayton, Victoria, Australia.
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Romana Stark
1Monash Biomedicine Discovery Institute and Department of Physiology, Monash University, Clayton, Victoria, Australia.
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Claire J. Foldi
1Monash Biomedicine Discovery Institute and Department of Physiology, Monash University, Clayton, Victoria, Australia.
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Philip Jean-Richard-dit-Bressel
2School of Psychology, UNSW, Sydney, Australia
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Zane B. Andrews
1Monash Biomedicine Discovery Institute and Department of Physiology, Monash University, Clayton, Victoria, Australia.
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  • For correspondence: Vasilios.Drakopoulos{at}monash.edu Zane.Andrews{at}monash.edu
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Abstract

Fiber photometry is a neuroscience technique that can continuously monitor in vivo fluorescence to assess population neural activity or neuropeptide/transmitter release in freely behaving animals. Despite the widespread adoption of this technique, methods to statistically analyse data in an unbiased, objective, and easily adopted manner are lacking. Various pipelines for data analysis exist, but they are often system-specific, only for pre-processing data, and/or lack usability. Current post hoc statistical approaches involve inadvertently biased user-defined time-binned averages or area under the curve analysis. To date, no post-hoc user-friendly tool with few assumptions for a standardised unbiased analysis exists, yet such a tool would improve reproducibility and statistical reliability for all users. Hence, we have developed a user-friendly post hoc statistical analysis package in Python that is easily downloaded and applied to data from any fiber photometry system. This Fiber Photometry Post Hoc Analysis (FiPhoPHA) package incorporates a variety of tools, a downsampler, bootstrapped confidence intervals (CIs) for analyzing peri-event signals between groups and compared to baseline, and permutation tests for comparing peri-event signals across comparison periods. We also include the ability to quickly and efficiently sort the data into mean time bins, if desired. This provides an open-source, user-friendly python package for unbiased and standardised post-hoc statistical analysis to improve reproducibility using data from any fiber photometry system.

Significance Statement Despite the widespread adoption of in vivo photometry for neuroscience research, methods to statistically analyse data in an unbiased, objective, and easily adopted manner are lacking. Various pipelines for data analysis exist, but they are often system-specific, only for pre-processing data, and/or lack usability. Current post hoc statistical approaches involve inadvertently biased user-defined time-binned averages or area under the curve analysis. Here, we have developed a standardised post hoc statistical analysis package in Python that is easily downloaded and applied to data from any fiber photometry system. This provides an open-source, user-friendly python package for unbiased and standardised post-hoc statistical analysis to improve reproducibility using data from any fiber photometry system.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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FiPhoPHA - A fiber photometry python package for post-hoc analysis
Vasilios Drakopoulos, Alex Reichenbach, Romana Stark, Claire J. Foldi, Philip Jean-Richard-dit-Bressel, Zane B. Andrews
eNeuro 29 July 2025, ENEURO.0221-25.2025; DOI: 10.1523/ENEURO.0221-25.2025

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FiPhoPHA - A fiber photometry python package for post-hoc analysis
Vasilios Drakopoulos, Alex Reichenbach, Romana Stark, Claire J. Foldi, Philip Jean-Richard-dit-Bressel, Zane B. Andrews
eNeuro 29 July 2025, ENEURO.0221-25.2025; DOI: 10.1523/ENEURO.0221-25.2025
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