Abstract

We compared performance of recently developed silicon photomultipliers (SiPMs) to GaAsP photomultiplier tubes (PMTs) for two-photon imaging of neural activity. Despite higher dark counts, SiPMs match or exceed the signal-to-noise ratio of PMTs at photon rates encountered in typical calcium imaging experiments due to their low pulse height variability. At higher photon rates encountered during high-speed voltage imaging, SiPMs substantially outperform PMTs.

© 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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2019 (5)

M. Castello, G. Tortarolo, M. Buttafava, T. Deguchi, F. Villa, S. Koho, L. Pesce, M. Oneto, S. Pelicci, L. Lanzanó, P. Bianchini, C. J. R. Sheppard, A. Diaspro, A. Tosi, and G. Vicidomini, “A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM,” Nat. Methods 16(2), 175–178 (2019).
[Crossref]

H. Dana, Y. Sun, B. Mohar, B. K. Hulse, A. M. Kerlin, J. P. Hasseman, G. Tsegaye, A. Tsang, A. Wong, R. Patel, J. J. Macklin, Y. Chen, A. Konnerth, V. Jayaraman, L. L. Looger, E. R. Schreiter, K. Svoboda, and D. S. Kim, “High-performance calcium sensors for imaging activity in neuronal populations and microcompartments,” Nat. Methods 16(7), 649–657 (2019).
[Crossref]

A. Kazemipour, O. Novak, D. Flickinger, J. S. Marvin, A. S. Abdelfattah, J. King, P. M. Borden, J. J. Kim, S. H. Al-Abdullatif, P. E. Deal, E. W. Miller, E. R. Schreiter, S. Druckmann, K. Svoboda, L. L. Looger, and K. Podgorski, “Kilohertz frame-rate two-photon tomography,” Nat. Methods 16(8), 778–786 (2019).
[Crossref]

R. Liu, Z. Li, J. S. Marvin, and D. Kleinfeld, “Direct wavefront sensing enables functional imaging of infragranular axons and spines,” Nat. Methods 16(7), 615–618 (2019).
[Crossref]

M. G. Giacomelli, “Evaluation of silicon photomultipliers for multiphoton and laser scanning microscopy,” J. Biomed. Opt. 24(10), 106503 (2019).
[Crossref]

2017 (3)

J. Wu, A. H. L. Tang, A. T. Y. Mok, W. Yan, G. C. F. Chan, K. K. Y. Wong, and K. K. Tsia, “Multi-MHz laser-scanning single-cell fluorescence microscopy by spatiotemporally encoded virtual source array,” Biomed. Opt. Express 8(9), 4160–4171 (2017).
[Crossref]

K. Wagatsuma, K. Miwa, M. Sakata, K. Oda, H. Ono, M. Kameyama, J. Toyohara, and K. Ishii, “Comparison between new-generation SiPM-based and conventional PMT-based TOF-PET/CT,” Physica Medica 42, 203–210 (2017).
[Crossref]

S. S. Kim, H. Rouault, S. Druckmann, and V. Jayaraman, “Ring attractor dynamics in the Drosophila central brain,” Science 356(6340), 849–853 (2017).
[Crossref]

2016 (5)

N. Li, K. Daie, K. Svoboda, and S. Druckmann, “Robust neuronal dynamics in premotor cortex during motor planning,” Nature 532(7600), 459–464 (2016).
[Crossref]

R. Prevedel, A. J. Verhoef, A. J. Pernia-Andrade, S. Weisenburger, B. S. Huang, T. Nöbauer, A. Fernández, J. E. Delcour, P. Golshani, A. Baltuska, and A. Vaziri, “Fast volumetric calcium imaging across multiple cortical layers using sculpted light,” Nat. Methods 13(12), 1021–1028 (2016).
[Crossref]

P. E. Deal, R. U. Kulkarni, S. H. Al-Abdullatif, and E. W. Miller, “Isomerically Pure Tetramethylrhodamine Voltage Reporters,” J. Am. Chem. Soc. 138(29), 9085–9088 (2016).
[Crossref]

A. Fukasawa, Y. Egawa, T. Ishizu, A. Kageyama, A. Kamiya, T. Muramatsu, G. Nakano, and Y. Negi, “Multichannel HPD for high-speed single photon counting,” Nucl. Instrum. Methods Phys. Res., Sect. A 812, 81–85 (2016).
[Crossref]

F. Mattioli, Z. Zhou, A. Gaggero, R. Gaudio, R. Leoni, and A. Fiore, “Photon-counting and analog operation of a 24-pixel photon number resolving detector based on superconducting nanowires,” Opt. Express 24(8), 9067–9076 (2016).
[Crossref]

2015 (5)

V. Emiliani, A. E. Cohen, K. Deisseroth, and M. Häusser, “All-Optical Interrogation of Neural Circuits,” J. Neurosci. 35(41), 13917–13926 (2015).
[Crossref]

A. Singh, J. D. McMullen, E. A. Doris, and W. R. Zipfel, “Comparison of objective lenses for multiphoton microscopy in turbid samples,” Biomed. Opt. Express 6(8), 3113–3127 (2015).
[Crossref]

J. W. Cha, E. Y. S. Yew, D. Kim, J. Subramanian, E. Nedivi, and P. T. C. So, “Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube,” AIP Adv. 5(8), 084802 (2015).
[Crossref]

D. Gudkov, G. Gudkov, B. Gorbovitski, and V. Gorfinkel, “Enhancing the Linear Dynamic Range in Multi-Channel Single Photon Detector Beyond 7OD,” IEEE Sens. J. 15(12), 7081–7086 (2015).
[Crossref]

T. Hige, Y. Aso, M. N. Modi, G. M. Rubin, and G. C. Turner, “Heterosynaptic Plasticity Underlies Aversive Olfactory Learning in Drosophila,” Neuron 88(5), 985–998 (2015).
[Crossref]

2013 (2)

R. Agishev, A. Comerón, J. Bach, A. Rodriguez, M. Sicard, J. Riu, and S. Royo, “Lidar with SiPM: Some capabilities and limitations in real environment,” Opt. Laser Technol. 49, 86–90 (2013).
[Crossref]

K. Podgorski and K. Haas, “Fast non-negative temporal deconvolution for laser scanning microscopy,” J. Biophotonics 6(2), 153–162 (2013).
[Crossref]

2012 (2)

M. Akiba, K. Inagaki, and K. Tsujino, “Photon number resolving SiPM detector with 1 GHz count rate,” Opt. Express 20(3), 2779–2788 (2012).
[Crossref]

S. Vinogradov, “Analytical models of probability distribution and excess noise factor of solid state photomultiplier signals with crosstalk,” Nucl. Instrum. Methods Phys. Res., Sect. A 695, 247–251 (2012).
[Crossref]

2011 (1)

J. D. Driscoll, A. Y. Shih, S. Iyengar, J. J. Field, G. A. White, J. A. Squier, G. Cauwenberghs, and D. Kleinfeld, “Photon counting, censor corrections, and lifetime imaging for improved detection in two-photon microscopy,” J. Neurophysiol. 105(6), 3106–3113 (2011).
[Crossref]

2010 (1)

E. A. Naumann, A. R. Kampff, D. A. Prober, A. F. Schier, and F. Engert, “Monitoring neural activity with bioluminescence during natural behavior,” Nat. Neurosci. 13(4), 513–520 (2010).
[Crossref]

2007 (4)

S. Tisa, A. Tosi, and F. Zappa, “Fully-integrated CMOS single photon counter,” Opt. Express 15(6), 2873–2887 (2007).
[Crossref]

E. Grigoriev, A. Akindinov, M. Breitenmoser, S. Buono, E. Charbon, C. Niclass, I. Desforges, and R. Rocca, “Silicon photomultipliers and their bio-medical applications,” Nucl. Instrum. Methods Phys. Res., Sect. A 571(1-2), 130–133 (2007).
[Crossref]

N. Dinu, R. Battiston, M. Boscardin, G. Collazuol, F. Corsi, G. F. Dalla Betta, A. Del Guerra, G. Llosá, M. Ionica, G. Levi, S. Marcatili, C. Marzocca, C. Piemonte, G. Pignatel, A. Pozza, L. Quadrani, C. Sbarra, and N. Zorzi, “Development of the first prototypes of Silicon PhotoMultiplier (SiPM) at ITC-irst,” Nucl. Instrum. Methods Phys. Res., Sect. A 572(1), 422–426 (2007).
[Crossref]

S. A. Castelletto, I. P. Degiovanni, V. Schettini, and A. L. Migdall, “Reduced deadtime and higher rate photon-counting detection using a multiplexed detector array,” J. Mod. Opt. 54(2-3), 337–352 (2007).
[Crossref]

2006 (1)

Z. Sadygov, A. Olshevski, I. Chirikov, I. Zheleznykh, and A. Novikov, “Three advanced designs of micro-pixel avalanche photodiodes: Their present status, maximum possibilities and limitations,” Nucl. Instrum. Methods Phys. Res., Sect. A 567(1), 70–73 (2006).
[Crossref]

2004 (1)

2003 (2)

P. Buzhan, B. Dolgoshein, L. Filatov, A. Ilyin, V. Kantzerov, V. Kaplin, A. Karakash, F. Kayumov, S. Klemin, E. Popova, and S. Smirnov, “Silicon photomultiplier and its possible applications,” Nucl. Instrum. Methods Phys. Res., Sect. A 504(1-3), 48–52 (2003).
[Crossref]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref]

2002 (1)

A. Bross, E. Flattum, D. Lincoln, S. Grünendahl, J. Warchol, M. Wayne, and P. Padley, “Characterization and performance of visible light photon counters (VLPCs) for the upgraded DØ detector at the Fermilab Tevatron,” Nucl. Instrum. Methods Phys. Res., Sect. A 477(1-3), 172–178 (2002).
[Crossref]

2000 (1)

E. Albrecht, M. Alemi, G. Barber, J. Bibby, M. Campbell, A. Duane, T. Gys, J. Montenegro, D. Piedigrossi, R. Schomaker, W. Snoeys, S. Wotton, and K. Wyllie, “Performance of hybrid photon detector prototypes with 80% active area for the rich counters of LHCB,” Nucl. Instrum. Methods Phys. Res., Sect. A 442(1-3), 164–170 (2000).
[Crossref]

1998 (1)

B. Cabrera, R. M. Clarke, P. Colling, A. J. Miller, S. Nam, and R. W. Romani, “Detection of single infrared, optical, and ultraviolet photons using superconducting transition edge sensors,” Appl. Phys. Lett. 73(6), 735–737 (1998).
[Crossref]

1996 (1)

1936 (1)

V. K. Zworykin, G. A. Morton, and L. Malter, “The Secondary Emission Multiplier-A New Electronic Device,” Proc. IRE 24(3), 351–375 (1936).
[Crossref]

Abdelfattah, A. S.

A. Kazemipour, O. Novak, D. Flickinger, J. S. Marvin, A. S. Abdelfattah, J. King, P. M. Borden, J. J. Kim, S. H. Al-Abdullatif, P. E. Deal, E. W. Miller, E. R. Schreiter, S. Druckmann, K. Svoboda, L. L. Looger, and K. Podgorski, “Kilohertz frame-rate two-photon tomography,” Nat. Methods 16(8), 778–786 (2019).
[Crossref]

Agishev, R.

R. Agishev, A. Comerón, J. Bach, A. Rodriguez, M. Sicard, J. Riu, and S. Royo, “Lidar with SiPM: Some capabilities and limitations in real environment,” Opt. Laser Technol. 49, 86–90 (2013).
[Crossref]

Akiba, M.

Akindinov, A.

E. Grigoriev, A. Akindinov, M. Breitenmoser, S. Buono, E. Charbon, C. Niclass, I. Desforges, and R. Rocca, “Silicon photomultipliers and their bio-medical applications,” Nucl. Instrum. Methods Phys. Res., Sect. A 571(1-2), 130–133 (2007).
[Crossref]

Al-Abdullatif, S. H.

A. Kazemipour, O. Novak, D. Flickinger, J. S. Marvin, A. S. Abdelfattah, J. King, P. M. Borden, J. J. Kim, S. H. Al-Abdullatif, P. E. Deal, E. W. Miller, E. R. Schreiter, S. Druckmann, K. Svoboda, L. L. Looger, and K. Podgorski, “Kilohertz frame-rate two-photon tomography,” Nat. Methods 16(8), 778–786 (2019).
[Crossref]

P. E. Deal, R. U. Kulkarni, S. H. Al-Abdullatif, and E. W. Miller, “Isomerically Pure Tetramethylrhodamine Voltage Reporters,” J. Am. Chem. Soc. 138(29), 9085–9088 (2016).
[Crossref]

Albrecht, E.

E. Albrecht, M. Alemi, G. Barber, J. Bibby, M. Campbell, A. Duane, T. Gys, J. Montenegro, D. Piedigrossi, R. Schomaker, W. Snoeys, S. Wotton, and K. Wyllie, “Performance of hybrid photon detector prototypes with 80% active area for the rich counters of LHCB,” Nucl. Instrum. Methods Phys. Res., Sect. A 442(1-3), 164–170 (2000).
[Crossref]

Alemi, M.

E. Albrecht, M. Alemi, G. Barber, J. Bibby, M. Campbell, A. Duane, T. Gys, J. Montenegro, D. Piedigrossi, R. Schomaker, W. Snoeys, S. Wotton, and K. Wyllie, “Performance of hybrid photon detector prototypes with 80% active area for the rich counters of LHCB,” Nucl. Instrum. Methods Phys. Res., Sect. A 442(1-3), 164–170 (2000).
[Crossref]

Aso, Y.

T. Hige, Y. Aso, M. N. Modi, G. M. Rubin, and G. C. Turner, “Heterosynaptic Plasticity Underlies Aversive Olfactory Learning in Drosophila,” Neuron 88(5), 985–998 (2015).
[Crossref]

Bach, J.

R. Agishev, A. Comerón, J. Bach, A. Rodriguez, M. Sicard, J. Riu, and S. Royo, “Lidar with SiPM: Some capabilities and limitations in real environment,” Opt. Laser Technol. 49, 86–90 (2013).
[Crossref]

Baltuska, A.

R. Prevedel, A. J. Verhoef, A. J. Pernia-Andrade, S. Weisenburger, B. S. Huang, T. Nöbauer, A. Fernández, J. E. Delcour, P. Golshani, A. Baltuska, and A. Vaziri, “Fast volumetric calcium imaging across multiple cortical layers using sculpted light,” Nat. Methods 13(12), 1021–1028 (2016).
[Crossref]

Barber, G.

E. Albrecht, M. Alemi, G. Barber, J. Bibby, M. Campbell, A. Duane, T. Gys, J. Montenegro, D. Piedigrossi, R. Schomaker, W. Snoeys, S. Wotton, and K. Wyllie, “Performance of hybrid photon detector prototypes with 80% active area for the rich counters of LHCB,” Nucl. Instrum. Methods Phys. Res., Sect. A 442(1-3), 164–170 (2000).
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Appl. Opt. (2)

Appl. Phys. Lett. (1)

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Biomed. Opt. Express (2)

IEEE Sens. J. (1)

D. Gudkov, G. Gudkov, B. Gorbovitski, and V. Gorfinkel, “Enhancing the Linear Dynamic Range in Multi-Channel Single Photon Detector Beyond 7OD,” IEEE Sens. J. 15(12), 7081–7086 (2015).
[Crossref]

J. Am. Chem. Soc. (1)

P. E. Deal, R. U. Kulkarni, S. H. Al-Abdullatif, and E. W. Miller, “Isomerically Pure Tetramethylrhodamine Voltage Reporters,” J. Am. Chem. Soc. 138(29), 9085–9088 (2016).
[Crossref]

J. Biomed. Opt. (1)

M. G. Giacomelli, “Evaluation of silicon photomultipliers for multiphoton and laser scanning microscopy,” J. Biomed. Opt. 24(10), 106503 (2019).
[Crossref]

J. Biophotonics (1)

K. Podgorski and K. Haas, “Fast non-negative temporal deconvolution for laser scanning microscopy,” J. Biophotonics 6(2), 153–162 (2013).
[Crossref]

J. Mod. Opt. (1)

S. A. Castelletto, I. P. Degiovanni, V. Schettini, and A. L. Migdall, “Reduced deadtime and higher rate photon-counting detection using a multiplexed detector array,” J. Mod. Opt. 54(2-3), 337–352 (2007).
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J. Neurophysiol. (1)

J. D. Driscoll, A. Y. Shih, S. Iyengar, J. J. Field, G. A. White, J. A. Squier, G. Cauwenberghs, and D. Kleinfeld, “Photon counting, censor corrections, and lifetime imaging for improved detection in two-photon microscopy,” J. Neurophysiol. 105(6), 3106–3113 (2011).
[Crossref]

J. Neurosci. (1)

V. Emiliani, A. E. Cohen, K. Deisseroth, and M. Häusser, “All-Optical Interrogation of Neural Circuits,” J. Neurosci. 35(41), 13917–13926 (2015).
[Crossref]

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref]

Nat. Methods (5)

M. Castello, G. Tortarolo, M. Buttafava, T. Deguchi, F. Villa, S. Koho, L. Pesce, M. Oneto, S. Pelicci, L. Lanzanó, P. Bianchini, C. J. R. Sheppard, A. Diaspro, A. Tosi, and G. Vicidomini, “A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM,” Nat. Methods 16(2), 175–178 (2019).
[Crossref]

R. Prevedel, A. J. Verhoef, A. J. Pernia-Andrade, S. Weisenburger, B. S. Huang, T. Nöbauer, A. Fernández, J. E. Delcour, P. Golshani, A. Baltuska, and A. Vaziri, “Fast volumetric calcium imaging across multiple cortical layers using sculpted light,” Nat. Methods 13(12), 1021–1028 (2016).
[Crossref]

A. Kazemipour, O. Novak, D. Flickinger, J. S. Marvin, A. S. Abdelfattah, J. King, P. M. Borden, J. J. Kim, S. H. Al-Abdullatif, P. E. Deal, E. W. Miller, E. R. Schreiter, S. Druckmann, K. Svoboda, L. L. Looger, and K. Podgorski, “Kilohertz frame-rate two-photon tomography,” Nat. Methods 16(8), 778–786 (2019).
[Crossref]

R. Liu, Z. Li, J. S. Marvin, and D. Kleinfeld, “Direct wavefront sensing enables functional imaging of infragranular axons and spines,” Nat. Methods 16(7), 615–618 (2019).
[Crossref]

H. Dana, Y. Sun, B. Mohar, B. K. Hulse, A. M. Kerlin, J. P. Hasseman, G. Tsegaye, A. Tsang, A. Wong, R. Patel, J. J. Macklin, Y. Chen, A. Konnerth, V. Jayaraman, L. L. Looger, E. R. Schreiter, K. Svoboda, and D. S. Kim, “High-performance calcium sensors for imaging activity in neuronal populations and microcompartments,” Nat. Methods 16(7), 649–657 (2019).
[Crossref]

Nat. Neurosci. (1)

E. A. Naumann, A. R. Kampff, D. A. Prober, A. F. Schier, and F. Engert, “Monitoring neural activity with bioluminescence during natural behavior,” Nat. Neurosci. 13(4), 513–520 (2010).
[Crossref]

Nature (1)

N. Li, K. Daie, K. Svoboda, and S. Druckmann, “Robust neuronal dynamics in premotor cortex during motor planning,” Nature 532(7600), 459–464 (2016).
[Crossref]

Neuron (1)

T. Hige, Y. Aso, M. N. Modi, G. M. Rubin, and G. C. Turner, “Heterosynaptic Plasticity Underlies Aversive Olfactory Learning in Drosophila,” Neuron 88(5), 985–998 (2015).
[Crossref]

Nucl. Instrum. Methods Phys. Res., Sect. A (8)

S. Vinogradov, “Analytical models of probability distribution and excess noise factor of solid state photomultiplier signals with crosstalk,” Nucl. Instrum. Methods Phys. Res., Sect. A 695, 247–251 (2012).
[Crossref]

E. Albrecht, M. Alemi, G. Barber, J. Bibby, M. Campbell, A. Duane, T. Gys, J. Montenegro, D. Piedigrossi, R. Schomaker, W. Snoeys, S. Wotton, and K. Wyllie, “Performance of hybrid photon detector prototypes with 80% active area for the rich counters of LHCB,” Nucl. Instrum. Methods Phys. Res., Sect. A 442(1-3), 164–170 (2000).
[Crossref]

A. Fukasawa, Y. Egawa, T. Ishizu, A. Kageyama, A. Kamiya, T. Muramatsu, G. Nakano, and Y. Negi, “Multichannel HPD for high-speed single photon counting,” Nucl. Instrum. Methods Phys. Res., Sect. A 812, 81–85 (2016).
[Crossref]

A. Bross, E. Flattum, D. Lincoln, S. Grünendahl, J. Warchol, M. Wayne, and P. Padley, “Characterization and performance of visible light photon counters (VLPCs) for the upgraded DØ detector at the Fermilab Tevatron,” Nucl. Instrum. Methods Phys. Res., Sect. A 477(1-3), 172–178 (2002).
[Crossref]

P. Buzhan, B. Dolgoshein, L. Filatov, A. Ilyin, V. Kantzerov, V. Kaplin, A. Karakash, F. Kayumov, S. Klemin, E. Popova, and S. Smirnov, “Silicon photomultiplier and its possible applications,” Nucl. Instrum. Methods Phys. Res., Sect. A 504(1-3), 48–52 (2003).
[Crossref]

E. Grigoriev, A. Akindinov, M. Breitenmoser, S. Buono, E. Charbon, C. Niclass, I. Desforges, and R. Rocca, “Silicon photomultipliers and their bio-medical applications,” Nucl. Instrum. Methods Phys. Res., Sect. A 571(1-2), 130–133 (2007).
[Crossref]

N. Dinu, R. Battiston, M. Boscardin, G. Collazuol, F. Corsi, G. F. Dalla Betta, A. Del Guerra, G. Llosá, M. Ionica, G. Levi, S. Marcatili, C. Marzocca, C. Piemonte, G. Pignatel, A. Pozza, L. Quadrani, C. Sbarra, and N. Zorzi, “Development of the first prototypes of Silicon PhotoMultiplier (SiPM) at ITC-irst,” Nucl. Instrum. Methods Phys. Res., Sect. A 572(1), 422–426 (2007).
[Crossref]

Z. Sadygov, A. Olshevski, I. Chirikov, I. Zheleznykh, and A. Novikov, “Three advanced designs of micro-pixel avalanche photodiodes: Their present status, maximum possibilities and limitations,” Nucl. Instrum. Methods Phys. Res., Sect. A 567(1), 70–73 (2006).
[Crossref]

Opt. Express (3)

Opt. Laser Technol. (1)

R. Agishev, A. Comerón, J. Bach, A. Rodriguez, M. Sicard, J. Riu, and S. Royo, “Lidar with SiPM: Some capabilities and limitations in real environment,” Opt. Laser Technol. 49, 86–90 (2013).
[Crossref]

Physica Medica (1)

K. Wagatsuma, K. Miwa, M. Sakata, K. Oda, H. Ono, M. Kameyama, J. Toyohara, and K. Ishii, “Comparison between new-generation SiPM-based and conventional PMT-based TOF-PET/CT,” Physica Medica 42, 203–210 (2017).
[Crossref]

Proc. IRE (1)

V. K. Zworykin, G. A. Morton, and L. Malter, “The Secondary Emission Multiplier-A New Electronic Device,” Proc. IRE 24(3), 351–375 (1936).
[Crossref]

Science (1)

S. S. Kim, H. Rouault, S. Druckmann, and V. Jayaraman, “Ring attractor dynamics in the Drosophila central brain,” Science 356(6340), 849–853 (2017).
[Crossref]

Other (2)

J. Rajchman and E. W. Pike, Electrostatic Focusing in Secondary Emission Multipliers. (RCA Manufacturing Co. RCA Victor Division Electronic Research Laboratory, 1937).

K. Yamamoto, K. Yamamura, K. Sato, T. Ota, H. Suzuki, and S. Ohsuka, “Development of Multi-Pixel Photon Counter (MPPC),” in 2006 IEEE Nuclear Science Symposium Conference Record (2006), 2, pp. 1094–1097.

Supplementary Material (1)

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» Visualization 1       Video 1

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Figures (3)

Fig. 1.
Fig. 1. PMT and SiPM operating principles. a. PMT operating principle. The photocathode converts a photon into a single electron with high efficiency. The electron is accelerated under high voltage, striking a series of dynodes. Each collision releases several more electrons, exponentially amplifying the signal. b. PMT gain variability. Simulated electron count distributions at different stages of PMT amplification. Stochastic variation in small integer numbers of electrons collected from the first dynode produces large variance in pulse heights, a form of multiplicative noise. Gain was modeled as Poisson. c. SiPM operating principle. An array of SPADs make up the SiPM. Each SPAD behaves like an ‘all or none’ switch, producing a stereotypical current pulse when one or more photons are absorbed. The output is the sum of the individual SPAD currents. Avalanches also occur without photon absorption, producing dark counts, a form of additive noise. d. SiPM gain variability. Simulated electron count distribution for a SiPM. Saturating amplification in each SPAD makes SiPM pulse heights highly uniform, with low multiplicative noise. e-f. Empirical pulse height spectra of a PMT (e) and SiPM (f), measured in the same sample simultaneously. g. Effects of additive and multiplicative noise on signal-to-noise ratio. See Equation (1). In these log-log plots, additive noise produces a signal-dependent offset, and multiplicative noise produces a constant offset. The relative effect of additive and multiplicative noise changes with photon rate. h. Measured SNR of PMT (blue) and SiPM (red) detectors during raster scanning at varying photon rates. Fit values for µ and α (Equation 1) are shown. Shaded region denotes photon rates encountered in raster scanning calcium imaging (Fig. 2(a-f)).
Fig. 2.
Fig. 2. Neuronal activity imaging with SiPM and PMT detectors. a-f. Simultaneous raster-scanning calcium imaging with a PMT and blue-sensitive SiPM using a 50:50 beamsplitter. a. Drosophila melanogaster olfactory pathway, mushroom body Kenyon cells expressing GCaMP6f (see Visualization 1). Image intensity is the square-root of the average of 300 frames. b. Simultaneously-measured ΔF/F0 traces for an example neuron. Gray bar indicates odor presentation period. c. SNR of simultaneously-recorded Ca2+- responses. Gray line denotes slope 1 diagonal. Paired two-sided t-test p < 1e-14. d. Mouse motor cortex imaged with a PMT and blue-sensitive SiPM using a 50:50 beamsplitter. Images are the square root of the average of 1000 frames. e. Simultaneously-measured ΔF/F0 traces for an example neuron. f. SNR of simultaneously-recorded Ca2+- responses. Gray line denotes slope 1 diagonal. Paired two-sided t-test p < 1e-10. g-i. High-speed voltage imaging using SiPMs and a PMT. g. Hippocampal neuron culture labeled with the red voltage-sensitive dye RhoVR1.pip.sulf. h. Neuron voltage traces (1016 Hz) recorded with a PMT and blue-sensitive SiPM (top, N = 3 neurons), or the blue-sensitive and peak-shifted SiPMs (bottom, N = 4 neurons). Each trace is a single trial. i. Mean action potential SNR for each voltage recording. Each neuron was recorded using two detectors, allowing within-neuron comparisons. *: p < 0.05, paired two-sided t-test
Fig. 3.
Fig. 3. Detector signals. a. Images acquired by our raster scanning microscope with no laser excitation, using the PMT (left) and blue-sensitive SiPM (right). Color maps are scaled to the maximum pixel brightness. Faint fixed pattern noise, with maximum amplitude less than a single photon, is seen for both detectors. Standard practices were used to minimize electromagnetic interference and avoid ground loops. We were unable to further reduce this noise in our labs. The SiPM showed larger fixed pattern noise relative to the single photon amplitude, contributing additional additive noise to raster-scanning measurements. SLAP measurements are not affected by this noise in the same way, due to how pixel intensities are computed (see b). b. PMT (top) and blue-sensitive SiPM (bottom) signals during imaging, recorded by the SLAP microscope digitizer. SiPM module signals were digitized at full bandwidth, with decay limited by passive quenching of Geiger-mode APDs. PMT signals were low-pass filtered by the amplifier at 40 MHz. Digitization is synchronized to the excitation laser clock. In the SLAP microscope, photon counts associated with each laser pulse are computed by local baseline-subtraction and regression against the single-photon response. c. Computed photon response for the blue-sensitive SiPM, as recorded by the SLAP microscope digitizer. Response is the median digitizer trace triggered on laser pulses while imaging a fluorescent sample at low power, scaled to the mean amplitude of single detection events.

Tables (1)

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Table 1. Measured quantum efficiency and dark rates of SiPM modules and GaAsP PMT

Equations (1)

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V a r ( x ^ ) = ( μ x ) + α

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