Deep Imaging in Scattering Media with Single Photon Selective Plane Illumination Microscopy (SPIM)

TL;DR

This paper demonstrates that Selective Plane Illumination Microscopy (SPIM) can achieve deep imaging in scattering tissues comparable to two-photon microscopy (2PM) without requiring high-powered lasers, offering a more accessible alternative.

Contribution

The study shows that SPIM can image nearly as deep as 2PM in scattering media, using theoretical and experimental methods, reducing the need for expensive pulsed lasers.

Findings

SPIM images more than twice as deep as epifluorescence and confocal microscopy.

SPIM reaches approximately 10 times the mean scattering length.

SPIM offers a cost-effective alternative for deep tissue imaging.

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. In this paper, using a combination of theoretical and experimental techniques, we show that 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 (approximately 10 times the mean scattering length). These results suggest that SPIM has the potential to provide deep imaging in scattering media in situations where 2PM systems would be too large or costly.