Scanless multitarget-matching multiphoton excitation fluorescence microscopy

Junpeng Qiu,Bruce Zhi Gao,Yonghong Shao,Lei Wang,Junle Qu

doi:10.1142/s1793545817500134

Junpeng Qiu, Bruce Zhi Gao + Show 3 more

Open Access

https://doi.org/10.1142/s1793545817500134

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Abstract

Using the combination of a reflective blazed grating and a reflective phase-only diffractive spatial light modulator (SLM), scanless multitarget-matching multiphoton excitation fluorescence microscopy (SMTM-MPM) was achieved. The SLM shaped an incoming mode-locked, near-infrared Ti:sapphire laser beam into an excitation pattern with addressable shapes and sizes that matched the samples of interest in the field of view. Temporal and spatial focusing were simultaneously realized by combining an objective lens and a blazed grating. The fluorescence signal from illuminated areas was recorded by a two-dimensional sCMOS camera. Compared with a conventional temporal focusing multiphoton microscope, our microscope achieved effective use of the laser power and decreased photodamage with higher axial resolution.

Highlights

As traditional multiphoton microscopy is based on single excitation beam scanning, imaging speed and the in°exible scanning method limit its applications in some important biological imaging processes
The temporal focusing system consisted of a tube lens and objective lens which executed temporal focusing to simultaneously excite the target samples of interest selected in wideeld illumination for each sub-region spot
We developed novel multitarget-matching multiphoton excitation °uorescence microscopy by combining spatial light modulator (SLM) and a grating

Summary

Introduction

Multiphoton excited °uorescence microscopy has become an important tool in theeld of biomedical optical imaging.[1,2,3] It is widely used in biomedical detection because nonlinear excitation of the nearinfrared femtosecond laser delivers low damage, a large depth of penetration in highly scattered tissues and high axial resolution with no confocal aperture.[4,5,6,7] As traditional multiphoton microscopy is based on single excitation beam scanning, imaging speed and the in°exible scanning method limit its applications in some important biological imaging processes. In the study of cardiomyocyte interaction with stem cells,[8] to avoid photodamage to cardiomyocytes during rapid imaging of the stem cell dynamic process, selective. This is an Open Access article published by World Scientic Publishing Company. Further distribution of this work is permitted, provided the original work is properly cited

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