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Fiber photometry is a significantly less invasive method compared to otherdeep brain imaging microendoscopy approaches due to the use of thin multimodefibers (MMF diameter $<$ 500 $\mu$m). Nevertheless, the transmitted signals getscrambled upon propagation within the MMF, thus limiting the technique'spotential in resolving temporal readouts with cellular resolution. Here, wedemonstrate how to separate the time trace signals of several fluorescentsources probed by a thin ($\approx$ 200 $\mu$m) MMF with typical implantablelength in a mouse brain. We disentangled several spatio-temporal fluorescencesignals by using a general unconstrained non-negative matrix factorization(NMF) algorithm directly on the raw video data. Furthermore, we show thatcommercial and low-cost open-source miniscopes display enough sensitivity toimage the same fluorescence patterns seen in our proof of principle experiment,suggesting that a new avenue for novel minimally invasive deep brain studieswith multimode fibers in freely-behaving mice could be possible.

Demixing fluorescence time traces transmitted by multimode fibers