CoChat
Sign Up
Cited by 47Advanced Fluorescence Microscopy TechniquesOptical Imaging and Spectroscopy TechniquesPhotochemistry and Electron Transfer Studies

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

Advancements in fluorescence lifetime imaging microscopy Instrumentation: Towards high speed and 3D

Open access
Park, Jongchan, Gao, Liang·2024·Current Opinion in Solid State and Materials Science
DOI 10.1016/j.cossms.2024.101147PMC PMC11290093
Open this paper in CoChat — free account

Abstract

Fluorescence lifetime imaging microscopy (FLIM) is a powerful imaging tool offering molecular specific insights into samples through the measurement of fluorescence decay time, with promising applications in diverse research fields. However, to acquire two-dimensional lifetime images, conventional FLIM relies on extensive scanning in both the spatial and temporal domain, resulting in much slower acquisition rates compared to intensity-based approaches. This problem is further magnified in three-dimensional imaging, as it necessitates additional scanning along the depth axis. Recent advancements have aimed to enhance the speed and three-dimensional imaging capabilities of FLIM. This review explores the progress made in addressing these challenges and discusses potential directions for future developments in FLIM instrumentation.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.cossms.2024.101147

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig. 1. Fluorescence lifetime imaging. (a) FLIM records time-resolved fluorescence decay at each spatial location. (b) Frequency-domain FLIM. (c) Time-domain FLIM.
Fig. 1. Fluorescence lifetime imaging. (a) FLIM records time-resolved fluorescence decay at each spatial location. (b) Frequency-domain FLIM. (c) Time-domain FLIM.CC BY-NC · PubMed Central
Fig. 2. High-speed FLIM methods. (a) The photon rate (throughput) of TCSPC-mode FLIM is limited to avoid measurement errors such as pile-up effects and counting loss. (b) The photon rate can be increased by using TCSPC with parallel detector arrays [22]. (c) Fluorescence decay waveform can be directly measured by high-speed photodetectors [23]. (d) Two-photon fluorescence lifetime dynamics of apoptosis induced cells were recorded at a video rate [24]. (e) Spectro-temporal encoded illumination enables kilohertz FLIM [25] and its application in (f) imaging flow cytometry.
Fig. 2. High-speed FLIM methods. (a) The photon rate (throughput) of TCSPC-mode FLIM is limited to avoid measurement errors such as pile-up effects and counting loss. (b) The photon rate can be increased by using TCSPC with parallel detector arrays [22]. (c) Fluorescence decay waveform can be directly measured by high-speed photodetectors [23]. (d) Two-photon fluorescence lifetime dynamics of apoptosis induced cells were recorded at a video rate [24]. (e) Spectro-temporal encoded illumination enables kilohertz FLIM [25] and its application in (f) imaging flow cytometry.CC BY-NC · PubMed Central
Fig. 3. High-speed FLIM methods using 2D detector arrays. (a) 0.5-megapixel SPAD camera with an acquisition rate up to 1 Hz [28] (b) 3.6 megapixel wide-field FLIM image captured by using the SPAD camera with image stitching [27]. (c) Time-folded FLIM. An optical cavity generates temporally delayed and spatially sheared replicas of fluorescent decay signal [29]. (d) The signal from the CMOS sensor and intensified CCD was used to generate fluorescence lifetime image with either an inverse retrieval method or neural network [29]. (e) Compressed ultrafast photography FLIM. The fluorescence signal is spatially mapped by the DMD, temporally sheared by the streak camera, and mapped onto the CCD sensor [31]. The fluorescence lifetime image is computationally reconstructed by solving the inverse problem. (f) High-speed fluorescence lifetime imaging of neural action potentials in live cells [30]. (g) Fluorescence lifetime imaging of neuronal cytoskeleton immunolabeled with multiple fluorophores.
Fig. 3. High-speed FLIM methods using 2D detector arrays. (a) 0.5-megapixel SPAD camera with an acquisition rate up to 1 Hz [28] (b) 3.6 megapixel wide-field FLIM image captured by using the SPAD camera with image stitching [27]. (c) Time-folded FLIM. An optical cavity generates temporally delayed and spatially sheared replicas of fluorescent decay signal [29]. (d) The signal from the CMOS sensor and intensified CCD was used to generate fluorescence lifetime image with either an inverse retrieval method or neural network [29]. (e) Compressed ultrafast photography FLIM. The fluorescence signal is spatially mapped by the DMD, temporally sheared by the streak camera, and mapped onto the CCD sensor [31]. The fluorescence lifetime image is computationally reconstructed by solving the inverse problem. (f) High-speed fluorescence lifetime imaging of neural action potentials in live cells [30]. (g) Fluorescence lifetime imaging of neuronal cytoskeleton immunolabeled with multiple fluorophores.CC BY-NC · PubMed Central
Fig. 4. 3D FLIM. (a) Light-sheet microscopy with a TCSPC-based wide-field FLIM detector [42]. (b) 3D volumetric fluorescence lifetime image of a mixture of fluorescent beads and quantum dots [42]. (c) 3D fluorescence lifetime imaging with optical projection tomography. A series of time-gated fluorescence images were acquired at various projection angles by rotating the sample [46]. (d) 3D volumetric fluorescence lifetime image of a live zebrafish [48]. (e) Light-field tomography FLIM. The 2D perspective images of a 3D sample are captured by pupil selection through a scanning mirror. The resultant images are further rotated and compressed along one spatial axis by a dove prism and cylindrical lens, respectively. (f) High-resolution depth-sectioned fluorescence lifetime images of a mouse kidney tissue section.
Fig. 4. 3D FLIM. (a) Light-sheet microscopy with a TCSPC-based wide-field FLIM detector [42]. (b) 3D volumetric fluorescence lifetime image of a mixture of fluorescent beads and quantum dots [42]. (c) 3D fluorescence lifetime imaging with optical projection tomography. A series of time-gated fluorescence images were acquired at various projection angles by rotating the sample [46]. (d) 3D volumetric fluorescence lifetime image of a live zebrafish [48]. (e) Light-field tomography FLIM. The 2D perspective images of a 3D sample are captured by pupil selection through a scanning mirror. The resultant images are further rotated and compressed along one spatial axis by a dove prism and cylindrical lens, respectively. (f) High-resolution depth-sectioned fluorescence lifetime images of a mouse kidney tissue section.CC BY-NC · PubMed Central
Fig. 5. Future perspective. (a) Compressed sensing method to overcome the information limit in FLIM [60]. (b) In vivo FLIM with a data compression ratio of 99% [59]. (c) 3D-stacked SPAD image sensor with a 45% fill factor [63]. (d) Fluorescence lifetime imaging of Convallaria Majalis sample at a video rate using the SPAD array [63]. (e) The mouse brain vasculature imaging through skin and skull. SWIR fluorescence enables deep tissue imaging beyond the NIR window [69]. (f) Lifetime characterization of SWIR fluorophores [70].
Fig. 5. Future perspective. (a) Compressed sensing method to overcome the information limit in FLIM [60]. (b) In vivo FLIM with a data compression ratio of 99% [59]. (c) 3D-stacked SPAD image sensor with a 45% fill factor [63]. (d) Fluorescence lifetime imaging of Convallaria Majalis sample at a video rate using the SPAD array [63]. (e) The mouse brain vasculature imaging through skin and skull. SWIR fluorescence enables deep tissue imaging beyond the NIR window [69]. (f) Lifetime characterization of SWIR fluorophores [70].CC BY-NC · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 70

  1. Berezin, 2010, Fluorescence lifetime measurements and biological imaging, Chem. Rev., 110, 2641, 10.1021/cr900343z · DOI
  2. Datta, 2020, Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications, J. Biomed. Opt., 25, 10.1117/1.JBO.25.7.071203 · DOI
  3. Alfonso-Garcia, 2021, Mesoscopic fluorescence lifetime imaging: fundamental principles, clinical applications and future directions, J. Biophotonics, 14, 10.1002/jbio.202000472 · DOI
  4. Dmitriev, 2021, Luminescence lifetime imaging of three-dimensional biological objects, J. Cell Sci., 134, 1, 10.1242/jcs.254763 · DOI
  5. Le Marois, 2017, Quantitative live cell FLIM imaging in three dimensions, Multi-Parametric Live Cell Microscopy of 3D Tissue Models, 31, 10.1007/978-3-319-67358-5_3 · DOI
  6. Becker, 2012, Fluorescence lifetime imaging – techniques and applications, J. Microsc., 247, 119, 10.1111/j.1365-2818.2012.03618.x · DOI · Paper page
  7. Borst, 2010, Fluorescence lifetime imaging microscopy in life sciences, Meas. Sci. Technol., 21, 10.1088/0957-0233/21/10/102002 · DOI · Paper page
  8. Ebrecht, 2014, Fluorescence lifetime imaging microscopy in the medical sciences, Protoplasma, 251, 293, 10.1007/s00709-013-0598-4 · DOI
  9. Marcu, 2012, Fluorescence lifetime techniques in medical applications, Ann. Biomed. Eng., 40, 304, 10.1007/s10439-011-0495-y · DOI
  10. Eichorst JP, Teng KW, Clegg RM: Fluorescence lifetime imaging techniques: Frequency-domain FLIM. In Fluorescence Lifetime Spectroscopy and Imaging: Principles and Applications in Biomedical Diagnostics. Edited by L Marcu PF, and DS Elson CRC Press; 2014:165-186.
  11. Becker W: Fluorescence lifetime imaging techniques: time-correlated single-photon counting. In Fluorescence Lifetime Spectroscopy and Imaging. Edited by: CRC Press; 2014:220-251. · DOI
  12. Gratton, 2003, Fluorescence lifetime imaging for the two-photon microscope: time-domain and frequency-domain methods, J. Biomed. Opt., 8, 381, 10.1117/1.1586704 · DOI
  13. Visscher, 1994, Fluorescence saturation in confocal microscopy, J. Microscopy-Oxford, 175, 162, 10.1111/j.1365-2818.1994.tb03479.x · DOI
  14. Liu, 2019, Fast fluorescence lifetime imaging techniques: a review on challenge and development, J. Innovative Optical Health Sci., 12, 1930003, 10.1142/S1793545819300039 · DOI
  15. Becker, 2019, Fast-acquisition TCSPC FLIM with sub-25-ps IRF width, Multiphoton Microscopy in the Biomedical Sci. XIX: SPIE, 10.1117/12.2510272 · DOI
  16. Liu, 2018, Parallelized fluorescence lifetime imaging microscopy (FLIM) based on photon reassignment, Opt. Commun., 421, 83, 10.1016/j.optcom.2018.03.060 · DOI
  17. Bower, 2018, High-speed imaging of transient metabolic dynamics using two-photon fluorescence lifetime imaging microscopy, Optica, 5, 1290, 10.1364/OPTICA.5.001290 · DOI
  18. Bower, 2019, Tracking metabolic dynamics of apoptosis with high-speed two-photon fluorescence lifetime imaging microscopy, Biomed. Opt. Express, 10, 6408, 10.1364/BOE.10.006408 · DOI
  19. Karpf, 2020, Spectro-temporal encoded multiphoton microscopy and fluorescence lifetime imaging at kilohertz frame-rates, Nat. Commun., 11, 2062, 10.1038/s41467-020-15618-w · DOI
  20. Bruschini, 2019, Single-photon avalanche diode imagers in biophotonics: review and outlook, Light Sci. Appl., 8, 87, 10.1038/s41377-019-0191-5 · DOI
  21. Zickus, 2020, Fluorescence lifetime imaging with a megapixel SPAD camera and neural network lifetime estimation, Sci. Rep., 10, 20986, 10.1038/s41598-020-77737-0 · DOI
  22. Morimoto, 2020, Megapixel time-gated SPAD image sensor for 2D and 3D imaging applications, Optica, 7, 346, 10.1364/OPTICA.386574 · DOI
  23. Kapitany, 2023, Single-shot time-folded fluorescence lifetime imaging, Proc. Natl. Acad. Sci., 120, 10.1073/pnas.2214617120 · DOI
  24. Ma, 2021, High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells, Proc. Natl. Acad. Sci., 118, 10.1073/pnas.2004176118 · DOI
  25. Gao, 2014, Single-shot compressed ultrafast photography at one hundred billion frames per second, Nature, 516, 74, 10.1038/nature14005 · DOI
  26. Liu, 2019, Single-shot compressed optical-streaking ultra-high-speed photography, Opt. Lett., 44, 1387, 10.1364/OL.44.001387 · DOI
  27. Liu, 2021, Fast wide-field upconversion luminescence lifetime thermometry enabled by single-shot compressed ultrahigh-speed imaging, Nat. Commun., 12, 6401, 10.1038/s41467-021-26701-1 · DOI
  28. Bowman, 2019, Electro-optic imaging enables efficient wide-field fluorescence lifetime microscopy, Nat. Commun., 10, 4561, 10.1038/s41467-019-12535-5 · DOI
  29. Bowman, 2021, Resonant electro-optic imaging for microscopy at Nanosecond resolution, ACS Nano, 15, 16043, 10.1021/acsnano.1c04470 · DOI
  30. Bowman, 2023, Wide-field fluorescence lifetime imaging of neuron spiking and subthreshold activity in vivo, Sci., 380, 1270, 10.1126/science.adf9725 · DOI
  31. Grant, 2005, Optically sectioned fluorescence lifetime imaging using a nipkow disk microscope and a tunable ultrafast continuum excitation source, Opt. Lett., 30, 3353, 10.1364/OL.30.003353 · DOI
  32. Grant, 2007, High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events, Opt. Express, 15, 15656, 10.1364/OE.15.015656 · DOI
  33. Mitchell, 2017, Functional in vivo imaging using fluorescence lifetime light-sheet microscopy, Opt. Lett., 42, 1269, 10.1364/OL.42.001269 · DOI
  34. Funane, 2018, Selective plane illumination microscopy (SPIM) with time-domain fluorescence lifetime imaging microscopy (FLIM) for volumetric measurement of cleared mouse brain samples, Rev. Sci. Instrum., 89, 10.1063/1.5018846 · DOI
  35. Hirvonen, 2020, Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting, J. Biophotonics, 13, 10.1002/jbio.201960099 · DOI
  36. Samimi K, Desa DE, Lin W, Weiss K, Li J, Huisken J, Miskolci V, Huttenlocher A, Chacko JV, Velten A, et al.: Light sheet autofluorescence lifetime imaging with a single photon avalanche diode array. bioRxiv 2023. · DOI
  37. Greger, 2011, Three-dimensional fluorescence lifetime imaging with a single plane illumination microscope provides an improved signal to noise ratio, Opt. Express, 19, 20743, 10.1364/OE.19.020743 · DOI
  38. Kumar AT: Tomographic fluorescence lifetime imaging. In Fluorescence Lifetime Spectroscopy and Imaging. Edited by: CRC Press; 2014:461-476.
  39. McGinty, 2008, Fluorescence lifetime optical projection tomography, J Biophotonics, 1, 390, 10.1002/jbio.200810044 · DOI
  40. McGinty, 2011, In vivo fluorescence lifetime optical projection tomography, Biomed. Opt. Express, 2, 1340, 10.1364/BOE.2.001340 · DOI
  41. Andrews, 2016, Visualising apoptosis in live zebrafish using fluorescence lifetime imaging with optical projection tomography to map FRET biosensor activity in space and time, J. Biophotonics, 9, 414, 10.1002/jbio.201500258 · DOI
  42. Sun, 2023, Optical projection tomography reconstruction with few views using highly-generalizable deep learning at sinogram domain, Biomed. Opt. Express, 14, 6260, 10.1364/BOE.500152 · DOI
  43. Ma, 2023, Light-field tomographic fluorescence lifetime imaging microscopy, PREPRINT at Res. Square
  44. Feng, 2021, Ultrafast light field tomography for snapshot transient and non-line-of-sight imaging, Nat. Commun., 12, 2179, 10.1038/s41467-021-22461-0 · DOI
  45. Feng, 2022, Compact light field photography towards versatile three-dimensional vision, Nat. Commun., 13, 3333, 10.1038/s41467-022-31087-9 · DOI
  46. Hirvonen, 2020, Fast timing techniques in FLIM applications, Front. Phys., 8, 161, 10.3389/fphy.2020.00161 · DOI
  47. Park, 2021, Review of bio-optical imaging systems with a high space-bandwidth product, Adv. Photonics, 3, 044001, 10.1117/1.AP.3.4.044001 · DOI
  48. Wetzstein, 2020, Inference in artificial intelligence with deep optics and photonics, Nature, 588, 39, 10.1038/s41586-020-2973-6 · DOI
  49. Tian, 2021, Deep Learning in biomedical optics, Lasers Surg. Med., 53, 748, 10.1002/lsm.23414 · DOI
  50. Brady, 2020, Deep learning for camera data acquisition, control, and image estimation, Adv. Opt. Photon., 12, 787, 10.1364/AOP.398263 · DOI
  51. Sorrells, 2021, Single-photon peak event detection (SPEED): a computational method for fast photon counting in fluorescence lifetime imaging microscopy, Opt. Express, 29, 37759, 10.1364/OE.439675 · DOI
  52. Ochoa, 2020, High compression deep learning based single-pixel hyperspectral macroscopic fluorescence lifetime imaging in vivo, Biomed. Opt. Express, 11, 5401, 10.1364/BOE.396771 · DOI
  53. Pian, 2017, Compressive hyperspectral time-resolved wide-field fluorescence lifetime imaging, Nat. Photonics, 11, 411, 10.1038/nphoton.2017.82 · DOI
  54. Lee, 2018, Progress in single-photon avalanche diode image sensors in standard CMOS: from two-dimensional monolithic to three-dimensional-stacked technology, Jpn. J. Appl. Phys., 10.7567/JJAP.57.1002A3 · DOI
  55. Hutchings, 2019, A reconfigurable 3-D-stacked SPAD imager with in-pixel histogramming for flash LIDAR or high-speed time-of-flight imaging, IEEE J. Solid State Circuits, 54, 2947, 10.1109/JSSC.2019.2939083 · DOI
  56. Erdogan, 2022, A high dynamic range 128 × 120 3-D stacked CMOS SPAD image sensor SoC for fluorescence microendoscopy, IEEE J. Solid State Circuits, 57, 1649, 10.1109/JSSC.2022.3150721 · DOI
  57. Matheson, 2023, Handheld wide-field fluorescence lifetime imaging system based on a distally mounted SPAD array, Opt. Express, 31, 22766, 10.1364/OE.482273 · DOI
  58. Choi, 2012, 3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation, Opt. Express, 20, 26219, 10.1364/OE.20.026219 · DOI
  59. Jia, 2021, Spatiotemporal control of biology: synthetic photochemistry toolbox with far-red and near-infrared light, ACS Chem. Biol., 17, 3255, 10.1021/acschembio.1c00518 · DOI
  60. Wong, 2022, Extending optical chemical tools and technologies to mice by shifting to the shortwave infrared region, Curr. Opin. Chem. Biol., 68, 10.1016/j.cbpa.2022.102131 · DOI · Paper page
  61. Cao, 2020, Recent progress in NIR-II contrast agent for biological imaging, Front. Bioeng. Biotechnol., 7, 487, 10.3389/fbioe.2019.00487 · DOI
  62. Carr, 2018, Shortwave infrared fluorescence imaging with the clinically approved near-infrared dye indocyanine green, Proc. Natl. Acad. Sci., 115, 4465, 10.1073/pnas.1718917115 · DOI · Paper page
  63. Chavez, 2023, Characterization of fluorescence lifetime of organic fluorophores for molecular imaging in the shortwave infrared window, J. Biomed. Opt., 28, 10.1117/1.JBO.28.9.094806 · DOI
  64. Thorburn, 2022, Ge-on-si single-photon avalanche diode detectors for short-wave infrared wavelengths, J. Phy: Photonics, 4
  65. Boas, 2001, Imaging the body with diffuse optical tomography, IEEE Signal Process Mag., 18, 57, 10.1109/79.962278 · DOI
  66. Kumar, 2008, A time domain fluorescence tomography system for small animal imaging, IEEE Trans. Med. Imaging, 27, 1152, 10.1109/TMI.2008.918341 · DOI
  67. Nothdurft, 2009, In vivo fluorescence lifetime tomography, J. Biomed. Opt., 10.1117/1.3086607 · DOI
  68. Chen, 2000, Optical computed tomography in a turbid medium using early arriving photons, J. Biomed. Opt., 5, 144, 10.1117/1.429981 · DOI
  69. Liu, 1993, Ultrafast laser-pulse transmission and imaging through biological tissues, Appl. Opt., 32, 554, 10.1364/AO.32.000554 · DOI
  70. Niedre, 2008, Early photon tomography allows fluorescence detection of lung carcinomas and disease progression in mice in vivo, Proc. Natl. Acad. Sci., 105, 19126, 10.1073/pnas.0804798105 · DOI

Cited by

Sorted by citations

See all 47

Metadata

Authors
Park, J., & Gao, L.
Affiliations
University of California, Los Angeles (US)
Corresponding author
Gao, Liang
ORCID
Venue
Current Opinion in Solid State and Materials Science
Publisher
Elsevier BV
Published
2024-06
Volume
30
Pages
101147
ISSN
1359-0286 (ISSN-L)
DOI
10.1016/j.cossms.2024.101147
PMC
PMC11290093
OpenAlex
W4392905680
Cited by
47
References
1
Citations by year
2026: 21 · 2025: 20 · 2024: 6
FWCI
2.54
Type
journal article
Language
en
Open access
Licence
Locations
  • Current Opinion in Solid State and Materials Science (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion, cc-by-nc)
Funders
  • National Institute of Neurological Disorders and Stroke (RF1NS128488)
  • National Institutes of Health
  • National Heart Lung and Blood Institute (R01HL165318)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
  • Physical Sciences › Chemistry › Physical and Theoretical Chemistry › Photochemistry and Electron Transfer Studies
Keywords
fluorescence lifetime imaging, 3D imaging, high-speed imaging, fluorescence lifetime, depth scanning

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review