A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging
Genetically encoded fluorescent calcium indicators allow cellular-resolution recording of physiology. However, bright, genetically targetable indicators that can be multiplexed with existing tools in vivo are needed for simultaneous imaging of multiple signals. Here we describe WHaloCaMP, a modular chemigenetic calcium indicator built from bright dye-ligands and protein sensor domains. Fluorescence change in WHaloCaMP results from reversible quenching of the bound dye via a strategically placed tryptophan. WHaloCaMP is compatible with rhodamine dye-ligands that fluoresce from green to near-infrared, including several that efficiently label the brain in animals. When bound to a near-infrared dye-ligand, WHaloCaMP shows a 7× increase in fluorescence intensity and a 2.1-ns increase in fluorescence lifetime upon calcium binding. We use WHaloCaMP1a to image Ca<sup>2+</sup> responses in vivo in flies and mice, to perform three-color multiplexed functional imaging of hundreds of neurons and astrocytes in zebrafish larvae and to quantify Ca<sup>2+</sup> concentration using fluorescence lifetime imaging microscopy (FLIM).
Authors
- Eric R. Schreiter (ORCID: https://orcid.org/0000-0002-2864-7469)
- Shang Fa Yang (ORCID: https://orcid.org/0000-0001-6671-7571)
- Yao Chen (ORCID: https://orcid.org/0000-0003-1509-6634)
- William C. Lemon (ORCID: https://orcid.org/0000-0003-4541-738X)
- Yajie Liang (ORCID: https://orcid.org/0000-0002-4798-4882)
- Christian Monroy Hernandez (ORCID: https://orcid.org/0000-0002-3796-989X)
- Helen Farrants (ORCID: https://orcid.org/0000-0003-2494-886X)
- Luke D. Lavis (ORCID: https://orcid.org/0000-0002-0789-6343)
- Carsen Stringer (ORCID: https://orcid.org/0000-0002-9229-4100)
- Guanda Qiao (ORCID: https://orcid.org/0000-0002-2483-6393)
- Michelle S. Frei (ORCID: https://orcid.org/0000-0002-4799-4554)
- Abraham G. Beyene (ORCID: https://orcid.org/0000-0003-3896-2144)
- Jonathan B. Grimm (ORCID: https://orcid.org/0000-0003-0331-4200)
- Ronak Patel (ORCID: https://orcid.org/0000-0002-6945-6466)
- Yichun Shuai (ORCID: https://orcid.org/0000-0001-9243-425X)
- Glenn Turner (ORCID: https://orcid.org/0000-0002-5341-2784)
- Filip Tomáška (ORCID: https://orcid.org/0000-0002-2546-9599)
- Philipp Keller (ORCID: https://orcid.org/0000-0003-2896-4920)
- Timothy Hanson (ORCID: https://orcid.org/0000-0002-6911-0094)
- Sarah E. Plutkis (ORCID: https://orcid.org/0009-0003-1822-0466)
- Deng Zhang (ORCID: https://orcid.org/0000-0002-1523-9503)
Institutions
- University of Maryland, Baltimore (US)
- Howard Hughes Medical Institute (US)
- University of California, Santa Barbara (US)
- Janelia Research Campus (US)
- Washington University in St. Louis (US)
- University of California San Diego (US)
- Max Planck Institute for Medical Research (DE)
Publication Details
- Journal
- Nature Methods
- Published
- 2024-09-20
- DOI
- https://doi.org/10.1038/s41592-024-02411-6
- Citations
- 46
- Primary Topic
- Neuroscience and Neuropharmacology Research
- Type
- article
- Field-Weighted Citation Impact
- 12.07
Funders
- Howard Hughes Medical Institute
- U.S. Department of Energy
- Maryland Stem Cell Research Fund
- Max-Planck-Gesellschaft
- Office of Science
- National Institute of Neurological Disorders and Stroke