Deep imaging in scattering media with selective plane illumination microscopy

Adithya Kumar Pediredla; Shizheng Zhang; Ben Avants; Fan Ye; Shin Nagayama; Ziying Chen; Caleb Kemere; Jacob T. Robinson; Ashok Veeraraghavan

doi:10.1117/1.JBO.21.12.126009

20 December 2016 Deep imaging in scattering media with selective plane illumination microscopy

Adithya Kumar Pediredla, Shizheng Zhang, Ben Avants, Fan Ye, Shin Nagayama, Ziying Chen, Caleb Kemere, Jacob T. Robinson, Ashok Veeraraghavan

Author Affiliations +

Journal of Biomedical Optics, Vol. 21, Issue 12, 126009 (December 2016). https://doi.org/10.1117/1.JBO.21.12.126009

Abstract

In most biological tissues, light scattering due to small differences in refractive index limits the depth of optical imaging systems. Two-photon microscopy (2PM), which significantly reduces the scattering of the excitation light, has emerged as the most common method to image deep within scattering biological tissue. This technique, however, requires high-power pulsed lasers that are both expensive and difficult to integrate into compact portable systems. Using a combination of theoretical and experimental techniques, we show that if the excitation path length can be minimized, selective plane illumination microscopy (SPIM) can image nearly as deep as 2PM without the need for a high-powered pulsed laser. Compared to other single-photon imaging techniques like epifluorescence and confocal microscopy, SPIM can image more than twice as deep in scattering media (∼10 times the mean scattering length). These results suggest that SPIM has the potential to provide deep imaging in scattering media in situations in which 2PM systems would be too large or costly.

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Adithya Kumar Pediredla, Shizheng Zhang, Ben Avants, Fan Ye, Shin Nagayama, Ziying Chen, Caleb Kemere, Jacob T. Robinson, and Ashok Veeraraghavan "Deep imaging in scattering media with selective plane illumination microscopy," Journal of Biomedical Optics 21(12), 126009 (20 December 2016). https://doi.org/10.1117/1.JBO.21.12.126009

Received: 26 August 2016; Accepted: 21 November 2016; Published: 20 December 2016

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