Abstract
Bioluminescence imaging (BLI) relies on the biochemical reaction between substrate and enzyme that triggers light emission upon convergence. Here, we present a protocol to study molecular oxygen dynamics in the in vivo mouse brain using the oxygen-dependent reaction between luciferase and its substrate. We describe steps for acute craniotomy, viral transfection, substrate administration, imaging, and analysis of hypoxic pockets. This protocol offers superior spatiotemporal properties compared to established approaches like electrodes and phosphorescence.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 103334 |
| Tidsskrift | STAR Protocols |
| Vol/bind | 5 |
| Udgave nummer | 4 |
| Antal sider | 23 |
| ISSN | 2666-1667 |
| DOI | |
| Status | Udgivet - 2024 |
Bibliografisk note
Funding Information:We would like to thank D. Xue for graphical support. This work was supported by the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (M.N.), National Institutes of Health grant R01AT012312 (M.N.), NINDS R01AT011439 (M.N.), U19 NS128613 (M.N.), the Simons Foundation (M.N.), Novo Nordisk Foundation NNF19OC0058058 (H.H.), Independent Research Fund Denmark 0134-00107B (H.H.), Novo Nordisk Foundation NNF20OC0066419 (M.N.), the Lundbeck Foundation R386-2021165 (M.N.), JPND/HBCI 1098-00030B (M.N.), JPND/Good Vibes 2092-00006B (M.N.), DOD W911NF2110006 (M.N.), the Lundbeck Foundation R287-2018-2046 (V.U.), Independent Research Fund Denmark 0134-00107B (H.H.), Independent Research Fund Denmark 3101-00282B (M.N.), US Army Research Office grant MURI W911NF1910280 (M.N.), Marie Sk\u0142odowska-Curie Fellowship ANCoDy 101064009 (A.A.), and an ONO Rising Star Fellowship (A.A.).
Funding Information:
We would like to thank D. Xue for graphical support. This work was supported by the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (M.N.), National Institutes of Health grant R01AT012312 (M.N.), NINDS R01AT011439 (M.N.), U19 NS128613 (M.N.), the Simons Foundation (M.N.), Novo Nordisk Foundation NNF19OC0058058 (H.H.), Independent Research Fund Denmark 0134-00107B (H.H.), Novo Nordisk Foundation NNF20OC0066419 (M.N.), the Lundbeck Foundation R386-2021165 (M.N.), JPND /HBCI 1098-00030B (M.N.), JPND /Good Vibes 2092-00006B (M.N.), DOD W911NF2110006 (M.N.), Independent Research Fund Denmark 3101-00282B (M.N.), US Army Research Office grant MURI W911NF1910280 (M.N.), Marie Sk\u0142odowska-Curie Fellowship ANCoDy 101064009 (A.A.), and an ONO Rising Star Fellowship (A.A.).
Publisher Copyright:
© 2024 The Author(s)