A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo

Simon Peter Poland; Grace Ka Yau Chan; James Alan Levitt; Nikola Krstajić; Ahmet Erdogan; Robert Henderson; Madeline Parsons; Simon Ameer-Beg

A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo

Simon Peter Poland, Grace Ka Yau Chan, James Alan Levitt, Nikola Krstajić, Ahmet Erdogan, Robert Henderson, Madeline Parsons, Simon Ameer-Beg

Research output: Contribution to journal › Letter › peer-review

Abstract

In this paper we will discuss the development of a multifocal multiphoton fluorescent lifetime imaging system where 4 individual fluorescent intensity and lifetime planes are acquired simultaneously, allowing us to obtain volumetric data without the need for sequential scanning at different axial depths. Using a phase-only SLM with an appropriate algorithm to generate a holographic pattern, we project a beamlet array within a sample volume of a size, which can be pre-programmed by the user. We demonstrate the capabilities of the system to image live-cell interactions. Whilst only 4 planes are shown, this technique can be rescaled to a large number of focal planes enabling full 3D acquisition and reconstruction.

Original language	English
Number of pages	4
Journal	Optics Letters
Publication status	Accepted/In press - 1 Nov 2018

Keywords

microscopy
FLUORESCENCE MICROSCOPY
nonlinear microscopy
detector arrays
diffractive optics
fluorescence lifetime imaging microscopy (FLIM)
FRET-FLIM

Cite this

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title = "A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo",

abstract = "In this paper we will discuss the development of a multifocal multiphoton fluorescent lifetime imaging system where 4 individual fluorescent intensity and lifetime planes are acquired simultaneously, allowing us to obtain volumetric data without the need for sequential scanning at different axial depths. Using a phase-only SLM with an appropriate algorithm to generate a holographic pattern, we project a beamlet array within a sample volume of a size, which can be pre-programmed by the user. We demonstrate the capabilities of the system to image live-cell interactions. Whilst only 4 planes are shown, this technique can be rescaled to a large number of focal planes enabling full 3D acquisition and reconstruction.",

keywords = "microscopy, FLUORESCENCE MICROSCOPY, nonlinear microscopy, detector arrays, diffractive optics, fluorescence lifetime imaging microscopy (FLIM), FRET-FLIM",

author = "Poland, {Simon Peter} and Chan, {Grace Ka Yau} and Levitt, {James Alan} and Nikola Krstaji{\'c} and Ahmet Erdogan and Robert Henderson and Madeline Parsons and Simon Ameer-Beg",

year = "2018",

month = nov,

day = "1",

language = "English",

journal = "Optics Letters",

issn = "0146-9592",

publisher = "The Optical Society",

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TY - JOUR

T1 - A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo

AU - Poland, Simon Peter

AU - Chan, Grace Ka Yau

AU - Levitt, James Alan

AU - Krstajić, Nikola

AU - Erdogan, Ahmet

AU - Henderson, Robert

AU - Parsons, Madeline

AU - Ameer-Beg, Simon

PY - 2018/11/1

Y1 - 2018/11/1

N2 - In this paper we will discuss the development of a multifocal multiphoton fluorescent lifetime imaging system where 4 individual fluorescent intensity and lifetime planes are acquired simultaneously, allowing us to obtain volumetric data without the need for sequential scanning at different axial depths. Using a phase-only SLM with an appropriate algorithm to generate a holographic pattern, we project a beamlet array within a sample volume of a size, which can be pre-programmed by the user. We demonstrate the capabilities of the system to image live-cell interactions. Whilst only 4 planes are shown, this technique can be rescaled to a large number of focal planes enabling full 3D acquisition and reconstruction.

AB - In this paper we will discuss the development of a multifocal multiphoton fluorescent lifetime imaging system where 4 individual fluorescent intensity and lifetime planes are acquired simultaneously, allowing us to obtain volumetric data without the need for sequential scanning at different axial depths. Using a phase-only SLM with an appropriate algorithm to generate a holographic pattern, we project a beamlet array within a sample volume of a size, which can be pre-programmed by the user. We demonstrate the capabilities of the system to image live-cell interactions. Whilst only 4 planes are shown, this technique can be rescaled to a large number of focal planes enabling full 3D acquisition and reconstruction.

KW - microscopy

KW - FLUORESCENCE MICROSCOPY

KW - nonlinear microscopy

KW - detector arrays

KW - diffractive optics

KW - fluorescence lifetime imaging microscopy (FLIM)

KW - FRET-FLIM

M3 - Letter

SN - 0146-9592

JO - Optics Letters

JF - Optics Letters

ER -

A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo

Abstract

Keywords

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