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Diffuse light-sheet microscopy for stripe-free calcium imaging of neural populations
Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review
DOI
- https://doi.org/10.1002/jbio.201800088
Final published version
- Michael A. Taylor, University of Queensland ,
- Gilles C. Vanwalleghem
- Itia A. Favre-Bulle, University of Queensland ,
- Ethan K. Scott, University of Queensland
Light-sheet microscopy is used extensively in developmental biology and neuroscience. One limitation of this approach is that absorption and scattering produces shadows in the illuminating light sheet, resulting in stripe artifacts. Here, we introduce diffuse light-sheet microscopes that use a line diffuser to randomize the light propagation within the image plane, allowing the light sheets to reform after obstacles. We incorporate diffuse light sheets in two existing configurations: selective plane illumination microscopy in which the sample is illuminated with a static sheet of light, and digitally scanned light sheet (DSLS) in which a thin Gaussian beam is scanned across the image plane during each acquisition. We compare diffuse light-sheet microscopes to their conventional counterparts for calcium imaging of neural activity in larval zebrafish. We show that stripe artifacts can cast deep shadows that conceal some neurons, and that the stripes can flicker, producing spurious signals that could be interpreted as biological activity. Diffuse light-sheets mitigate these problems, illuminating the blind spots produced by stripes and removing artifacts produced by the stripes' movements. The upgrade to diffuse light sheets is simple and inexpensive, especially in the case of DSLS, where it requires the addition of one optical element.
| Original language | English |
|---|---|
| Article number | e201800088 |
| Journal | journal of biophotonics |
| Volume | 11 |
| Issue | 12 |
| ISSN | 1864-063X |
| DOIs | |
| Publication status | Published - Dec 2018 |
| Externally published | Yes |
Bibliographical note
Funding Information:
information Australian Research Council, Grant/Award Numbers: DP140102036, DP110103612, FT110100887; European Molecular Biology Organization, Grant/Award Number: Long-term fellowship; Human Frontier Science Program, Grant/Award Number: LT000146/2016; National Health and Medical Research Council, Grant/Award Number: APP1066887; Simons Foundation, Grant/Award Number: Pilot Award (019851)We thank Harrison Wood for help assembling the solenoid actuator in the dSPIM experiment and Shaun Walters assistance with the synthetic fluorescent medium. Support was provided by the Australian National Fabrication Facility (ANFF), QLD node. Support was provided by the an NHMRC Project Grant (APP1066887), ARC Future Fellowship (FT110100887), a Simons Foundation Pilot Award (019851), and two ARC Discovery Project Grants (DP140102036 & DP110103612) to E.K.S.; an EMBO Long-term Fellowship to G.C.V.; and a fellowship from the Human Frontiers Science Program (LT000146/2016) to M.A.T.
Funding Information:
Australian Research Council, Grant/Award Numbers: DP140102036, DP110103612, FT110100887; European Molecular Biology Organization, Grant/Award Number: Long-term fellowship; Human Frontier Science Program, Grant/Award Number: LT000146/2016; National Health and Medical Research Council, Grant/ Award Number: APP1066887; Simons Foundation, Grant/Award Number: Pilot Award (019851)
Funding Information:
We thank Harrison Wood for help assembling the solenoid actuator in the dSPIM experiment and Shaun Walters assistance with the synthetic fluorescent medium. Support was provided by the Australian National Fabrication Facility (ANFF), QLD node. Support was provided by the an NHMRC Project Grant (APP1066887), ARC Future Fellowship (FT110100887), a Simons Foundation Pilot Award (019851), and two ARC Discovery Project Grants (DP140102036 & DP110103612) to E.K.S.; an EMBO Long-term Fellowship to G.C.V.; and a fellowship from the Human Frontiers Science Program (LT000146/2016) to M.A.T.
Publisher Copyright:
© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
- calcium imaging, fluorescence, light-sheet microscope, zebrafish
Research areas
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