CaltechAUTHORS
  A Caltech Library Service

High-speed, long-term, 4D in vivo lifetime imaging in intact and injured zebrafish and mouse brains by instant FLIM

Zhang, Yide and Guldner, Ian H. and Nichols, Evan L. and Benirschke, David and Smith, Cody J. and Zhang, Siyuan and Howard, Scott S. (2020) High-speed, long-term, 4D in vivo lifetime imaging in intact and injured zebrafish and mouse brains by instant FLIM. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20200207-102104833

[img] PDF (August 12, 2020) - Submitted Version
See Usage Policy.

9Mb
[img] Video (QuickTime) (Supplementary Video 1 - 3D slices of fixed mouse brain) - Supplemental Material
See Usage Policy.

28Mb
[img] Video (QuickTime) (Supplementary Video 2 - 3D GSOS slices of fixed mouse brain) - Supplemental Material
See Usage Policy.

3576Kb
[img] Video (QuickTime) (Supplementary Video 3 - Living mouse brain imaged with 3D instant FLIM with AO) - Supplemental Material
See Usage Policy.

12Mb
[img] Video (QuickTime) (Supplementary Video 4 - 4D reconstruction of zebrafish with laser injury) - Supplemental Material
See Usage Policy.

6Mb
[img] Video (QuickTime) (Supplementary Video 5 - 4D reconstruction of mouse brain with laser injury) - Supplemental Material
See Usage Policy.

4Mb
[img] Video (QuickTime) (Supplementary Video 6 - Summary of challenging mouse brain image) - Supplemental Material
See Usage Policy.

3360Kb
[img] Video (QuickTime) (Supplementary Video 7 - Phasor ROI labeling for 4D instant FLIM data) - Supplemental Material
See Usage Policy.

4Mb
[img] Video (QuickTime) (Supplementary Video 8 - Phasor K-means clustering for 4D instant FLIM data) - Supplemental Material
See Usage Policy.

3177Kb
[img] Video (QuickTime) (Supplementary Video 9 - Qualitative surface rendering of 4D FLIM of zebrafish) - Supplemental Material
See Usage Policy.

2974Kb

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20200207-102104833

Abstract

Traditional fluorescence microscopy is blind to molecular microenvironment information that is present in the emission decay lifetime. With fluorescence lifetime imaging microscopy (FLIM), physiological parameters such as pH, refractive index, ion concentration, dissolved gas concentration, and fluorescence resonance energy transfer (FRET) can be measured. Despite these benefits, existing FLIM techniques are typically slow, noisy, and hard to implement due to expensive instrumentation and complex post-processing. To overcome these limitations, we present instant FLIM, a method that allows real-time acquisition and display of two-photon intensity, lifetime, and phasor imaging data. Using analog signal processing, we demonstrate in vivo four-dimensional (4D) FLIM movies by imaging mouse and zebrafish glial cell response to injury over 12 hours through intact skulls. Instant FLIM can be implemented as an upgrade to an existing multiphoton microscope using cost-effective off-the-shelf components, requires no data post-processing, and is demonstrated to be compatible with FD-FLIM super-resolution techniques.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/2020.02.05.936039DOIDiscussion Paper
https://github.com/yzhang34/Instant-FLIM-Control.gitRelated ItemData/Code
https://github.com/yzhang34/Instant-FLIM-Analysis.gitRelated ItemData/Code
ORCID:
AuthorORCID
Zhang, Yide0000-0002-9463-3970
Guldner, Ian H.0000-0001-8715-136X
Nichols, Evan L.0000-0001-9835-9107
Benirschke, David0000-0002-9943-177X
Smith, Cody J.0000-0002-9831-1514
Zhang, Siyuan0000-0003-0910-3666
Howard, Scott S.0000-0003-3246-6799
Alternate Title:Instant FLIM enables 4D in vivo lifetime imaging of intact brains
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. This work was supported by the University of Notre Dame, the Center for Zebrafish Research and Center of Stem Cells and Regenerative Medicine at the University of Notre Dame, the National Science Foundation under Grant No. CBET-1554516 (S.S.H.), the National Institutes of Health R01 CA194697, R01 CA222405 (S.Z.), R01 NS107553 (C.J.S), the Elizabeth and Michael Gallagher Family (C.J.S.), the Alfred P. Sloan Foundation (C.J.S.), and the Berry Family Foundation Graduate Fellowship of Advanced Diagnostics & Therapeutics (AD&T) (Y.Z.). Author contributions: Y.Z. and S.S.H. conceived and coordinated the project, designed and built the microscope, wrote the control and image-processing software, and performed the imaging experiments. I.H.G. and S.Z. performed the mice surgery, acquired the images, and analyzed the data. E.L.N. and C.J.S. prepared the zebrafish, performed the zebrafish imaging experiments, and analyzed the data. D.B. assisted with the microscope assembly and performed the temperature-dependent imaging experiment. S.S.H., S.Z., and C.J.S. supervised the study. All authors contributed to the writing and editing of the manuscript. The authors declare no competing financial interests.
Funders:
Funding AgencyGrant Number
University of Notre DameUNSPECIFIED
NSFCBET-1554516
NIHR01 CA194697
NIHR01 CA222405
NIHR01 NS107553
Elizabeth and Michael Gallagher FamilyUNSPECIFIED
Alfred P. Sloan FoundationUNSPECIFIED
Berry Family FoundationUNSPECIFIED
Record Number:CaltechAUTHORS:20200207-102104833
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200207-102104833
Official Citation:High-speed, long-term, 4D in vivo lifetime imaging in intact and injured zebrafish and mouse brains by instant FLIM. Yide Zhang, Ian H. Guldner, Evan L. Nichols, David Benirschke, Cody J. Smith, Siyuan Zhang, Scott S. Howard. bioRxiv 2020.02.05.936039; doi: https://doi.org/10.1101/2020.02.05.936039
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:101180
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:10 Feb 2020 15:30
Last Modified:14 Aug 2020 16:03

Repository Staff Only: item control page