Two-dimensional random access multiphoton spatial frequency modulated imaging.

Alyssa M Allende Motz,Charles Durfee,Jeff Field,Randy Bartels,Christopher L Hoy,Daniel E Adams,Jeff Squier,John Czerski

doi:10.1364/oe.378460

Abstract

Spatial frequency modulated imaging (SPIFI) enables the use of an extended excitation source for linear and nonlinear imaging with single element detection. To date, SPIFI has only been used with fixed excitation source geometries. Here, we explore the potential for the SPIFI method when a spatial light modulator (SLM) is used to program the excitation source, opening the door to a more versatile, random access imaging environment. In addition, an in-line, quantitative pulse compensation and measurement scheme is demonstrated using a new technique, spectral phase and amplitude retrieval and compensation (SPARC). This enables full characterization of the light exposure conditions at the focal plane of the random access imaging system, an important metric for optimizing, and reporting imaging conditions within specimens.

Highlights

Multiphoton microscopy (MPM) [1,2,3,4,5,6,7,8,9,10] is an important subset of emergent biomedical imaging techniques
Such qualities make the MPM family of techniques highly suited for biomolecular research and have allowed MPM systems to become the tool of choice for the study of live cellular-level dynamics deep within scattering tissue specimens [3,4,5]
A series of measurements demonstrates the capabilities of the two-dimensional, Random Access Multiphoton or RAMP-Spatial frequency modulated imaging (SPIFI) method over a broad range of specimen types, imaging modalities and applications

Summary

Introduction

Multiphoton microscopy (MPM) [1,2,3,4,5,6,7,8,9,10] is an important subset of emergent biomedical imaging techniques. For a typical MPM system, operating with a near infrared (NIR) source in the 700-1300 nm range, this amounts to lateral spatial resolution at or below the micrometer-scale, at up to millimeter-scale depths within specimens [6,7]. Such qualities make the MPM family of techniques highly suited for biomolecular research and have allowed MPM systems to become the tool of choice for the study of live cellular-level dynamics deep within scattering tissue specimens [3,4,5]. MPM techniques have found use in several adjacent areas of research besides microbiology and biomedicine, such as characterization of materials [17,18], or as a probe for phenomena such as exciton lifetime [19,20]

Methods

Results

Conclusion