Fluorescence lifetime images of different green fluorescent proteins in fly brain

Sih-Yu Lai; Y. Y. Lin; A. S. Chiang; Y. C. Huang

doi:10.1117/12.808974

23 February 2009 Fluorescence lifetime images of different green fluorescent proteins in fly brain

Sih-Yu Lai, Y. Y. Lin, A. S. Chiang, Y. C. Huang

Proceedings Volume 7180, Photons and Neurons; 71800D (2009) https://doi.org/10.1117/12.808974
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

The mechanisms of learning and memory are the most important functions in an animal brain. Investigating neuron circuits and network maps in a brain is the first step toward understanding memory and learning behavior. Since Drosophila brain is the major model for understanding brain functions, we measure the florescence lifetimes of different GFP-based reporters expressed in a fly brain. In this work, two Gal4 drivers, OK 107 and MZ 19 were used. Intracellular calcium ([Ca²⁺]) concentration is an importation indicator of neuronal activity. Therefore, several groups have developed GFP-based calcium sensors, among which G-CaMP is the most popular and reliable. The fluorescence intensity of G-CaMP will increase when it binds to calcium ion; however, individual variation from different animals prevents quantitative research. In this work, we found that the florescence lifetime of G-CaMP will shrink from 1.8 ns to 1.0 ns when binding to Ca²⁺. This finding can potentially help us to understand the neuron circuits by fluorescence lifetime imaging microscopy (FLIM). Channelrhodopsin-2 (ChR2) is a light-activated ion-channel protein on a neuron cell membrane. In this work, we express ChR2 and G-CaMP in a fly brain. Using a pulsed 470-nm laser to activate the neurons, we can also record the fluorescence lifetime changes in the structure. Hence, we can trace and manipulate a specific circuit in this animal. This method provides more flexibility in brain research.

Citation Download Citation

Sih-Yu Lai, Y. Y. Lin, A. S. Chiang, and Y. C. Huang "Fluorescence lifetime images of different green fluorescent proteins in fly brain", Proc. SPIE 7180, Photons and Neurons, 71800D (23 February 2009); https://doi.org/10.1117/12.808974

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