PUBLICATION
Light-sheet autofluorescence lifetime imaging with a single-photon avalanche diode array
- Authors
- Samimi, K., Desa, D.E., Lin, W., Weiss, K., Li, J., Huisken, J., Miskolci, V., Huttenlocher, A., Chacko, J.V., Velten, A., Rogers, J.D., Eliceiri, K.W., Skala, M.C.
- ID
- ZDB-PUB-230624-36
- Date
- 2023
- Source
- Journal of Biomedical Optics 28: 066502066502 (Journal)
- Registered Authors
- Huttenlocher, Anna
- Keywords
- autofluorescence, fluorescence lifetime imaging microscopy, light-sheet microscopy, nicotinamide adenine dinucleotide (phosphate), single-photon avalanche diode
- MeSH Terms
-
- Animals
- Microscopy, Fluorescence/methods
- NAD*/metabolism
- Optical Imaging/methods
- Pancreatic Neoplasms*
- Photons
- Zebrafish
- PubMed
- 37351197 Full text @ J. Biomed. Opt.
Citation
Samimi, K., Desa, D.E., Lin, W., Weiss, K., Li, J., Huisken, J., Miskolci, V., Huttenlocher, A., Chacko, J.V., Velten, A., Rogers, J.D., Eliceiri, K.W., Skala, M.C. (2023) Light-sheet autofluorescence lifetime imaging with a single-photon avalanche diode array. Journal of Biomedical Optics. 28:066502066502.
Abstract
Significance Fluorescence lifetime imaging microscopy (FLIM) of the metabolic co-enzyme nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] is a popular method to monitor single-cell metabolism within unperturbed, living 3D systems. However, FLIM of NAD(P)H has not been performed in a light-sheet geometry, which is advantageous for rapid imaging of cells within live 3D samples.
Aim We aim to design, validate, and demonstrate a proof-of-concept light-sheet system for NAD(P)H FLIM.
Approach A single-photon avalanche diode camera was integrated into a light-sheet microscope to achieve optical sectioning and limit out-of-focus contributions for NAD(P)H FLIM of single cells.
Results An NAD(P)H light-sheet FLIM system was built and validated with fluorescence lifetime standards and with time-course imaging of metabolic perturbations in pancreas cancer cells with 10 s integration times. NAD(P)H light-sheet FLIM in vivo was demonstrated with live neutrophil imaging in a larval zebrafish tail wound also with 10 s integration times. Finally, the theoretical and practical imaging speeds for NAD(P)H FLIM were compared across laser scanning and light-sheet geometries, indicating a 30 × 6 ×
Conclusions FLIM of NAD(P)H is feasible in a light-sheet geometry and is attractive for 3D live cell imaging applications, such as monitoring immune cell metabolism and migration within an organism.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping