A pragmatic guide to multiphoton microscope design

Young, M.G.; Field, Jeffrey J.; Sheetz, Kraig E.; Bartels, Randy A.; Squier, Jeff

doi:10.1364/aop.7.000276

Cited by 52 publications

(26 citation statements)

References 331 publications

(362 reference statements)

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“…Photons per pixel can be estimated from parameters including the gain and offset of the photomultiplier tube (http://labrigger.com/blog/2010/07/30/measuring-the-gain-of-your-imaging-system/). Tradeoffs between photons per pixel, numbers of pixels, and SNR can quantified using signal detection theory, and compared and optimized for specific experiments 99, 100101 . For example, GCaMP6s imaging can provide single AP detection with nearly 100% detection of all spikes when imaged at a high frame rate over a small field-of-view (30 μm × 30 μm at 60 frames/s) 91 .…”

Section: Population Recording Via Calcium Imagingmentioning

confidence: 99%

Improving data quality in neuronal population recordings

et al. 2016

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Understanding how the brain operates requires understanding how large sets of neurons function together. Modern recording technology makes it possible to simultaneously record the activity of hundreds of neurons, and technological developments will soon allow recording of thousands or tens of thousands. As with all experimental techniques, these methods are subject to confounds that complicate the interpretation of such recordings, and could lead to erroneous scientific conclusions. Here, we discuss methods for assessing and improving the quality of data from these techniques, and outline likely future directions in this field.

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Section: Population Recording Via Calcium Imagingmentioning

confidence: 99%

Improving data quality in neuronal population recordings

et al. 2016

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“…In principle, the resolution of 2PM and 3PM could reach 1 √ 2 and 1 √ 3 times, respectively, of the 1PM resolution, because their signal intensities depend quadratically or cubically on the input light intensity [29]. However, this is usually difficult to achieve in experiment due to aberrations [12,[30][31][32]. As shown by Zemax simulation in Fig.…”

Section: Performance Characterizationmentioning

confidence: 99%

Miniaturized multimodal multiphoton microscope for simultaneous two-photon and three-photon imaging with a dual-wavelength Er-doped fiber laser

Huang

Zhou

Liu

et al. 2020

Biomed. Opt. Express

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A multimodal multiphoton microscopy (MPM) is developed to acquire both twophoton microscopy (2PM) and three-photon microscopy (3PM) signals. A dual-wavelength Er-doped fiber laser is used as the light source, which provides the fundamental pulse at 1580 nm to excite third harmonic generation (THG) and the frequency-doubled pulse at 790 nm to excite intrinsic two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG). Due to their different contrast mechanisms, the TPEF, SHG, and THG images can acquire complementary information about tissues, including cells, collagen fibers, lipids, and interfaces, all label-free. The compact MPM imaging probe is developed using miniature objective lens and a micro-electro-mechanical scanner. Furthermore, the femtosecond laser pulses are delivered by a single mode fiber and the signals are collected by a multimode fiber, which makes the miniaturized MPM directly fiber-coupled, compact, and portable. Design considerations on using the dual excitation wavelengths are discussed. Multimodal and label-free imaging by TPEF, SHG, and THG are demonstrated on biological samples. The miniaturized multimodal MPM is shown to have great potential for label-free imaging of thick and live tissues.

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“…Imaging modalities such as Stimulated Raman (SRS) and Coherent anti-Stokes Raman spectroscopy (CARS) require utilization of multiple wavelengths at the same time and at the same spatial location. It is possible to minimize the effects of chromatic aberration and dispersion in a multiphoton microscope system by various methods [21,22]. However, it is desirable to be able to remove this concern entirely.…”

Section: Introductionmentioning

confidence: 99%

All-reflective multiphoton microscope

et al. 2017

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Abstract:We present the design, construction, and characterization of a multiphoton microscope that uses reflective elements for beam shaping and steering. This compact all reflective design removes the adverse effects of dispersion on laser pulse broadening as well as chromatic aberration in the focusing of broadband and multicolored laser sources. The design of this system is discussed in detail, including aberrations analysis via ray-tracing simulation and opto-mechanical design. The resolution of this mirror based all-reflective microscope is characterized using fluorescent microbeads. The performance of the system at multiple wavelengths is investigated along with some potential multiphoton imaging and writing applications. Nelson, and S. Pané, "Hybrid Helical Magnetic Microrobots Obtained by 3D Template-Assisted Electrodeposition," Small 10(7), 1284-1288 (2014). 15. T. Gissibl, S. Thiele, A. Herkommer, and H. Giessen, "Two-photon direct laser writing of ultracompact multilens objectives," Nat. Photonics 10(8), 554-560 (2016). 73-76 (1990). 17. E. E. Hoover and J. A. Squier, "Advances in multiphoton microscopy technology," Nat. Photonics 7(2), 93-101 (2013). 18. K. Kieu, S. Mehravar, R. Gowda, R. A. Norwood, and N. Peyghambarian, "Label-free multi-photon imaging using a compact femtosecond fiber laser mode-locked by carbon nanotube saturable absorber," Biomed. Opt. Express 4(10), 2187-2195 (2013). 19. N. G. Horton and C. Xu, "Dispersion compensation in three-photon fluorescence microscopy at 1,700 nm,"Biomed. Opt. Express 6(4), 1392-1397 (2015). 20. I. D. Tullis, S. M. Ameer-Beg, P. R. Barber, V. Rankov, and B. Vojnovic, "Mapping femtosecond pulse front distortion and group velocity dispersion in multiphoton microscopy," Proc. SPIE 6089, 60890Y (2006). 21. W. Wang, Y. Liu, P. Xi, and Q. Ren, "Origin and effect of high-order dispersion in ultrashort pulse multiphoton microscopy in the 10 fs regime," Appl. Opt. 49(35), 6703-6709 (2010). 22. M. D. Young, J. J. Field, K. E. Sheetz, R. A. Bartels, and J. Squier, "A pragmatic guide to multiphoton microscope design," Adv. Opt. Photonics 7(2), 276-378 (2015). 23. J. Y. Yu, C. S. Liao, Z. Y. Zhuo, C. H. Huang, H. C. Chui, and S. W. Chu, "A diffraction-limited scanning system providing broad spectral range for laser scanning microscopy," Rev.

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A pragmatic guide to multiphoton microscope design

Cited by 52 publications

References 331 publications

Improving data quality in neuronal population recordings

Improving data quality in neuronal population recordings

Miniaturized multimodal multiphoton microscope for simultaneous two-photon and three-photon imaging with a dual-wavelength Er-doped fiber laser

All-reflective multiphoton microscope

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