Stanford
Deisseroth Lab

FIP

Frame-projected Independent-fiber Photometry (FIP)

Instructions for setting up Frame-projected Independent-fiber Photometry (FIP), also known as 'multi-fiber photometry' (MFP), as seen in Nature Methods:

Kim, Christina K., Samuel J. Yang, Nandini Pichamoorthy, Noah P. Young, Isaac Kauvar, Joshua H. Jennings, Talia N. Lerner et al. "Simultaneous fast measurement of circuit dynamics at multiple sites across the mammalian brain." Nature methods (2016).

To order parts and set up hardware, see Hardware.

Once hardware is set up, to configure software (written in Matlab), see Software.

 

Hardware

Download FIP Parts List

Ordering instructions:

  • Order parts from each vendor: Thorlabs (short lead time), Semrock (~1 week lead time), Point Grey (short lead time if in stock), National Instruments (short lead time if in stock), Doric Lenses (longest lead time), Amazon (short lead time if Prime)
  • Thorlabs rapid order file: download this and upload to here

Set up and alignment:

Here is the general schematic, as found in the paper:

Here is a labeled photograph of the completed setup:

Alignment instructions:

  • Lay out all pieces on the MB1224 first
  • Insert dichroic into DFM1, ensure that the dichroic is oriented in the correct way (check chroma or semrock website for their convention)
  • Connect cubes with ERSCA, then mount to breadboard using posts and forks
  • Connect everything else
  • Position fiber bundle tip at working distance of the objective. Ensure that LED illumination is as uniform as possible through all fibers of the bundle. Ensure that camera is focused onto the end of the fiber bundle.
  • Hook up LED drivers to nidaq or microcontroller for controlling alternating illumination

Software

Ensure you have Matlab (tested with >2013b) installed, with the data acquisition and image acquisition toolboxes.

Clone or download package from: https://github.com/deisseroth-lab/multifiber

For lightweight, do-it-yourself software: the only functions really necessary are the ability to record camera frames and to control the illumination. Then, post processing consists of selecting an ROI in within each fiber of the bundle and extracting the time series.