We have successfully demonstrated video-rate CMOS single-photon avalanche diode (SPAD)-based cameras for fluorescence lifetime imaging microscopy (FLIM) by applying innovative FLIM algorithms. We also review and compare several time-domain techniques and solid-state FLIM systems, and adapt the proposed algorithms for massive CMOS SPAD-based arrays and hardware implementations. The theoretical error equations are derived and their performances are demonstrated on the data obtained from 0.13 μm CMOS SPAD arrays and the multiple-decay data obtained from scanning PMT systems. In vivo two photon fluorescence lifetime imaging data of FITC-albumin labeled vasculature of a P22 rat carcinosarcoma (BD9 rat window chamber) are used to test how different algorithms perform on bi-decay data. The proposed techniques are capable of producing lifetime images with enough contrast.
References
[1]
Medintz, I.L.; Uyeda, H.T.; Goldman, E.R.; Mattoussi, H. Quantum dot bioconjugates for imaging, labelling and sensing. Nat. Mater. 2005, 4, 435–446.
[2]
Carlin, L.M.; Evans, R.; Milewicz, H.; Matthews, D.R.; Fernandes, L.; Perani, M.; Levitt, J.; Keppler, M.; Monypenny, J.; Coolen, A.; et al. A targeted siRNA screen identifies regulators of Cdc42 activity at the Natural Killer cell immunological synapse. Sci. Signal. 2011, doi:10.1126/scisignal.2001729.
[3]
Fruhwirth, G.O.; Fernandes, L.P.; Weitsman, G.; Patel, G.; Kelleher, M.; Lawler, K.; Brock, A.; Poland, S.P.; Mattews, D.R.; Keri, G.; et al. How f？ster resonance energy transfer imaging improves the understanding of protein interaction networks in cancer biology. Chem. Phys. Chem. 2011, 12, 442–461.
[4]
Lakowicz, J.R.; Szmacinski, H.; Nowaczyk, K.; Johnson, M.L. Fluorescence lifetime imaging of calcium using Quin-2. Cell Calcium 1992, 13, 131–147.
[5]
Spring, B.Q.; Clegg, R.M. Image analysis for denoising full-field frequency-domain fluorescence lifetime images. J. Microsc. 2009, 235, 221–237.
[6]
Gratton, E.; Breusegem, S.; Sutin, J.; Ruan, Q.; Barry, N. Fluorescence lifetime imaging for the two-photon microscope: Time-domain and frequency-domain methods. J. Biomed. Opt. 2003, 8, 381.
[7]
Clayton, A.H.A.; Hanley, Q.S.; Verveer, P.J. Graphical representation and multicomponent analysis of single-frequency fluorescence lifetime imaging microscopy data. J. Microsc. 2004, 213, 1–5.
[8]
Becker, W. Advanced Time-Correlated Single Photon Counting Techniques; Springer: New York, NY, USA, 2005.
[9]
Yoon, H.-J.; Itoh, S.; Kawahito, S. A CMOS image sensor with in-pixel two-stage charge transfer for fluorescence lifetime imaging. IEEE Trans. Electron. Devices 2009, 56, 214–221.
[10]
Huang, T.-C.D.; Sorgenfrei, S.; Gong, P.; Levicky, R.; Shepard, K.L. A 0.18 μm CMOS array sensor for integrated time-resolved fluorescence detection. IEEE J. Solid-State Circuits 2009, 44, 1644–1654.
[11]
Pancheri, L.; Stoppa, D. A SPAD-Based Pixel Linear Array for High-Speed Time-Gated Fluorescence Lifetime Imaging. Proceedings of the 34th IEEE European Solid-State Circuits Conference (ESSCIRC), Athens, Greece, 14–18 September 2009; pp. 428–431.
[12]
Benetti, M.; Iori, D.; Pancheri, L.; Borghetti, F.; Pasquardini, L.; Lunelli, L.; Pederzolli, C.; Gonzo, L.; Betta, G.-F.D.; Stoppa, D. Highly parallel SPAD detector for time-resolved lab-on-chip. Proc. SPIE 2010, 7723, 77231Q.
Li, D.-U.; Arlt, J.; Richardson, J.; Walker, R.; Buts, A.; Stoppa, D.; Charbon, E.; Henderson, R. Real-time fluorescence lifetime imaging system with a 32 × 32 0.13 μm CMOS low dark-count single-photon avalanche diode array. Opt. Express 2010, 18, 10257–10269.
[15]
Li, D.D.-U.; Arlt, J.; Tyndall, D.; Walker, R.; Richardson, J.; Stoppa, D.; Charbon, E.; Henderson, R.K. Video-rate fluorescence lifetime imaging camera with CMOS single-photon avalanche diode arrays and high-speed imaging algorithm. J. Biomed. Opt. 2011, 16, 096012.
[16]
Veerappan, C.; Richardson, J.; Walker, R.; Li, D.-U.; Fishburn, M.W.; Maruyama, Y.; Stoppa, D.; Borghetti, F.; Gersbach, M.; Henderson, R.K.; et al. A 160 × 128 Single-Photon Image Sensor with on-pixel 55ps 10bit Time-to-Digital Converter. Proceedings of the IEEE International Solid-State Circuits Conference (ISSCC) Dig. Tech. Papers, San Francisco, CA, USA, 20–24 February, 2011; pp. 312–314.
[17]
Maruyama, Y.; Charbon, E. An All-Digital, Time-Gated 128 × 128 SPAD Array for On-Chip, Filter-Less Fluorescence Detection. Proceedings of the 16th International Solid-State Sensors, Actuators and Microsystems Conference (Transducers), Beijing, China, 5–9 June 2011; pp. 1180–1183.
[18]
Guo, J.; Sonkusale, S. A CMOS Imager with Digital Phase Readout for Fluorescence Lifetime Imaging. Proceedings of the 36th IEEE European Solid-State Circuits Conference (ESSCIRC), Helsinki, Finland, 12–16 September 2011; pp. 115–118.
[19]
Yao, L.; Yung, K.Y.; Cheung, M.C.; Chodavarapu, V.P.; Bright, F.V. CMOS Direct Time Interval Measurement of Long-Lived Luminescence Lifetimes. Proceedings of the 33rd Annual International Conference of the IEEE EMBS, Boston, MA, USA, 30 August–3 September 2011; pp. 5–9.
[20]
Tyndall, D.; Rae, B.; Li, D.; Richardson, J.; Arlt, J.; Henderson, R. A 100Mphoton/s Time-Resolved Mini-Silicon Photomultiplier with On-Chip Fluorescence Lifetime Estimation in 0.13 μm CMOS Imaging Technology. Proceedings of the IEEE Int. Solid-State Circuits Conference (ISSCC) Dig. Tech. Papers, San Francisco, CA, USA, 19–23 February 2012; pp. 122–123.
[21]
Ballew, R.M.; Demas, J.N. An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays. Anal. Chem. 1989, 61, 30–33.
[22]
Richardson, J.; Walker, R.; Grant, L.; Stoppa, D.; Borghetti, F.; Charbon, E.; Gersbach, M.; Henderson, R. A 32 × 32 50ps Resolution 10bit Time to Digital Converter Array in 130 nm CMOS for Time Correlated Imaging. Proceedings of the IEEE Custom Integrated Circuits Conference (CICC), San Jose, CA, USA, 13–16 September 2009; pp. 77–80.
[23]
Li, L.-Q.; Davis, L.M. Single photon avalanche diode for single molecule detection. Rev. Sci. Instrum. 1993, 64, 1524–1529.
Niclass, C.; Soga, M. A Miniature Actively Recharged Single-Photon Detector Free of Afterpulsing Effects with 6 ns Dead Time in a 0.18 μm CMOS Technology. Proceedings of the IEEE International Electron Devices Meeting (IEDM) 2010, San Francisco, CA, USA, 6–8 December 2010; pp. 14.3:1–14.3:4.
[27]
Richardson, J.A.; Grant, L.A.; Henderson, R.K. Low dark count single-photon avalanche diode structure compatible with standard nanometer scale CMOS technology. IEEE Photon. Tech. Lett. 2009, 21, 1020–1022.
[28]
Henderson, R.K.; Webster, E.A.G.; Walker, R.; Richardson, J.A.; Grant, L.A. A 3 × 3, 5 μm Pitch, 3-Transistor Single Photon Avalanche Diode Array with Integrated 11 V Bias Generation in 90 nm CMOS Technology. Proceedings of the 2010 IEEE International Electron Devices Meeting (IEDM), San Francisco, CA, USA, 6–8 December 2010; pp. 14.2:1–14.2:4.
Webster, E.A.G.; Richardson, J.A.; Grant, L.A.; Renshaw, D.; Henderson, R.K. A single-photon avalanche diode in 90-nm CMOS imaging technology with 44% photon detection efficiency at 690 nm. IEEE Electron. Dev. Lett. 2012, 33, 694–696.
[31]
Niclass, C.; Soga, M.; Matsubara, H.; Kato, S. A 100 m-Range 10-Frame/s 340 × 96-Pixel Time-of-Flight Depth Sensor in 0.18 μm CMOS. Proceedings of the 36th IEEE European Solid-State Circuits Conference (ESSCIRC), Helsinki, Finland, 12–16 September 2011; pp. 107–110.
[32]
Elangovan, M.; Day, R.N.; Periasamy, A. Nanosecond fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy to localize the protein interactions in a single living cell. J. Microsc. 2002, 205, 3–14.
[33]
Sharman, K.K.; Periasamy, A. Error analysis of the rapid lifetime determination method for double-exponential decays and new windowing schemes. Anal. Chem. 1999, 71, 947–952.
Li, D.-U.; Bonnist Renshaw, D.; Henderson, R. On-chip time-correlated fluorescence lifetime extraction algorithms and error analysis. J. Opt. Soc. Am. A 2008, 25, 1190–1198.
[43]
Li, D.-U.; Andrews, R.; Arlt, J.; Henderson, R. Hardware implementation algorithm and error analysis of high-speed fluorescence lifetime sensing systems using center-of-mass method. J. Biomed. Opt. 2010, 15, 017006.
[44]
Draaijer, A.; Sanders, R.; Gerritsen, H.C. Fluorescence Lifetime Imaging, a New Tool in Confocal Microscopy. In Handbook of Biological Confocal Microscopy; Pawley, J., Ed.; Plenum Publishing: New York, NY, USA, 1995; pp. 491–505.
[45]
Hall, P.; Selinger, B. Better estimates of exponential decay parameters. J. Phys. Chem. 1981, 85, 2941–2946.
[46]
Li, D.-U.; Walker, R.; Richardson, J.; Rae, B.; Buts, A.; Renshaw, D.; Henderson, R. Hardware implementation and calibration of background noise for an integration-based fluorescence lifetime sensing algorithm. J. Opt. Soc. Am. 2009, 26, 804–814.
Moon, S; Won, Y.J.; Kim, D.Y. Analog mean-delay method for high-speed fluorescence lifetime measurement. Opt. Express 2009, 17, 2834–2849.
[49]
Wong, Y.J.; Han, W.-T.; Kim, D.Y. Precision and accuracy of the analog mean-delay method for high-speed fluorescence lifetime measurement. J. Opt. Soc. Am. A 2011, 28, 2026–2032.
[50]
Won, Y.J.; Moon, S.; Yang, W.B.; Kim, D.G.; Han, W.-T.; Kim, D.Y. High-speed confocal fluorescence lifetime imaging microscopy (FLIM) with the analog mean delay (AMD) method. Opt. Express 2011, 19, 3396–3405.
[51]
Padilla-Parra, S.; Auduge, N.; Coppey-Moisan, M.; Tramier, M. Quantitative FRET analysis by fast acquisition time domain FLIM at high spatial resolution in living cells. Biophys. J. 2008, 95, 2976–2988.
[52]
Yamada, H.; Padilla-Parra, S.; Park, S.-J.; Itoh, T.; Chaineau, M.; Monaldi, I.; Cremona, O.; Benfenati, F.; Camilli, P.D.; Coppey-Moisan, M.; et al. Dynamic interaction of amphiphysin with N-WASP regulates actin assembly. J. Biol. Chem. 2009, 284, 34244–34256.
[53]
Barber, P.R.; Ameer-Beg, S.M.; Pathmannanthan, S.; Rowly, M.; Collen, A.C.C. A Bayesian method for single molecule fluorescence burst analysis. Biomed. Opt. Express 2010, 1, 1148–1158.
[54]
Rowley, M.I; Barber, P.R.; Coolen, A.C.C.; Vojnovic, B. Bayesian analysis of fluorescence lifetime imaging data. Proc. SPIE 2011, 7903, 790325:1–790325:9.
[55]
Ameer-Big, S.M.; Barber, P.R.; Hodgkiss, R.J.; Locke, R.J.; Newman, R.G.; Tozer, G.M.; Vojnovic, B.; Wilson, J. Application of multiphoton steady state and lifetime imaging to mapping of tumour vascular architecture. in vivo. Proc. SPIE 2002, 4620, 85–95.
[56]
Papenfuss, H.D.; Gross, J.F.; Intaglietta, M.; Treese, F.A. A transparent access chamber for the rat dorsal skin fold. Microvasc. Res. 1979, 18, 311–318.
[57]
Dewhirst, M.W.; Gustafson, C.; Gross, J.F.; Tso, C.Y. Temporal effects of 5.0 Gy radiation in healing subcutaneous micro-vasculature of a dorsal flap window chamber. Rad. Res. 1987, 112, 581–591.
