A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919

D R Matthews; S M Ameer-Beg; P Barber; G P Pierce; R G Newman; B Vojnovic; L M Carlin; M D Keppler; T Ng; K Suhling; M Irving; D L Farkas; D V Nicolau; R C Leif

A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919

D R Matthews, S M Ameer-Beg, P Barber, G P Pierce, R G Newman, B Vojnovic, L M Carlin, M D Keppler, T Ng, K Suhling, M Irving, D L Farkas (Editor), D V Nicolau (Editor), R C Leif (Editor)

Research output: Chapter in Book/Report/Conference proceeding › Conference paper

Abstract

An automated high-content screening microscope has been developed which uses fluorescence anisotropy imaging and fluorescence lifetime microscopy to identify Forster resonant energy transfer between eGFP and mRPF1 in drug screening assays. A wide-field polarization resolved imager is used to simultaneously capture the parallel and perpendicular components of both eGFP and mRFP1 fluorescence emission to provide a high-speed measurement of acceptor depolarization. Donor excited state lifetime measurements performed using laser scanning microscopy is then used to determine the FRET efficiency in a particular assay. A proof-of-principle assay is performed using mutant Jurkat human T-cells to illustrate the process by which FRET is first identified and then quantified by our high-content screening system

Original language	English
Title of host publication	Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi
Place of Publication	BELLINGHAM
Publisher	SPIE - INT SOC OPTICAL ENGINEERING
Pages	85919 - 85919
Number of pages	1
ISBN (Print)	0277-786X
Publication status	Published - 2008
Event	Conference on Imaging, Manipulation and Analysis of Biomolecules, Cells, and Tissues VI - San Jose, CA Duration: 1 Jan 2008 → …

Publication series

Name	PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)

Conference

Conference	Conference on Imaging, Manipulation and Analysis of Biomolecules, Cells, and Tissues VI
City	San Jose, CA
Period	1/01/2008 → …

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Matthews, D. R., Ameer-Beg, S. M., Barber, P., Pierce, G. P., Newman, R. G., Vojnovic, B., Carlin, L. M., Keppler, M. D., Ng, T., Suhling, K., Irving, M., Farkas, D. L. (Ed.), Nicolau, D. V. (Ed.), & Leif, R. C. (Ed.) (2008). A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919. In Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi (pp. 85919 - 85919). (PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)). SPIE - INT SOC OPTICAL ENGINEERING.

Matthews, D R ; Ameer-Beg, S M ; Barber, P et al. / A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919. Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi. BELLINGHAM : SPIE - INT SOC OPTICAL ENGINEERING, 2008. pp. 85919 - 85919 (PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)).

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title = "A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919",

abstract = "An automated high-content screening microscope has been developed which uses fluorescence anisotropy imaging and fluorescence lifetime microscopy to identify Forster resonant energy transfer between eGFP and mRPF1 in drug screening assays. A wide-field polarization resolved imager is used to simultaneously capture the parallel and perpendicular components of both eGFP and mRFP1 fluorescence emission to provide a high-speed measurement of acceptor depolarization. Donor excited state lifetime measurements performed using laser scanning microscopy is then used to determine the FRET efficiency in a particular assay. A proof-of-principle assay is performed using mutant Jurkat human T-cells to illustrate the process by which FRET is first identified and then quantified by our high-content screening system",

author = "Matthews, {D R} and Ameer-Beg, {S M} and P Barber and Pierce, {G P} and Newman, {R G} and B Vojnovic and Carlin, {L M} and Keppler, {M D} and T Ng and K Suhling and M Irving and Farkas, {D L} and Nicolau, {D V} and Leif, {R C}",

year = "2008",

language = "English",

isbn = "0277-786X",

series = "PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)",

publisher = "SPIE - INT SOC OPTICAL ENGINEERING",

pages = "85919 -- 85919",

booktitle = "Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi",

note = "Conference on Imaging, Manipulation and Analysis of Biomolecules, Cells, and Tissues VI ; Conference date: 01-01-2008",

}

Matthews, DR, Ameer-Beg, SM, Barber, P, Pierce, GP, Newman, RG, Vojnovic, B , Carlin, LM, Keppler, MD, Ng, T , Suhling, K , Irving, M, Farkas, DL (ed.), Nicolau, DV (ed.) & Leif, RC (ed.) 2008, A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919. in Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi. PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE), SPIE - INT SOC OPTICAL ENGINEERING, BELLINGHAM, pp. 85919 - 85919, Conference on Imaging, Manipulation and Analysis of Biomolecules, Cells, and Tissues VI, San Jose, CA, 1/01/2008.

A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919. / Matthews, D R; Ameer-Beg, S M; Barber, P et al.
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi. BELLINGHAM: SPIE - INT SOC OPTICAL ENGINEERING, 2008. p. 85919 - 85919 (PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)).

Research output: Chapter in Book/Report/Conference proceeding › Conference paper

TY - CHAP

T1 - A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919

AU - Matthews, D R

AU - Ameer-Beg, S M

AU - Barber, P

AU - Pierce, G P

AU - Newman, R G

AU - Vojnovic, B

AU - Carlin, L M

AU - Keppler, M D

AU - Ng, T

AU - Suhling, K

AU - Irving, M

A2 - Farkas, D L

A2 - Nicolau, D V

A2 - Leif, R C

PY - 2008

Y1 - 2008

N2 - An automated high-content screening microscope has been developed which uses fluorescence anisotropy imaging and fluorescence lifetime microscopy to identify Forster resonant energy transfer between eGFP and mRPF1 in drug screening assays. A wide-field polarization resolved imager is used to simultaneously capture the parallel and perpendicular components of both eGFP and mRFP1 fluorescence emission to provide a high-speed measurement of acceptor depolarization. Donor excited state lifetime measurements performed using laser scanning microscopy is then used to determine the FRET efficiency in a particular assay. A proof-of-principle assay is performed using mutant Jurkat human T-cells to illustrate the process by which FRET is first identified and then quantified by our high-content screening system

AB - An automated high-content screening microscope has been developed which uses fluorescence anisotropy imaging and fluorescence lifetime microscopy to identify Forster resonant energy transfer between eGFP and mRPF1 in drug screening assays. A wide-field polarization resolved imager is used to simultaneously capture the parallel and perpendicular components of both eGFP and mRFP1 fluorescence emission to provide a high-speed measurement of acceptor depolarization. Donor excited state lifetime measurements performed using laser scanning microscopy is then used to determine the FRET efficiency in a particular assay. A proof-of-principle assay is performed using mutant Jurkat human T-cells to illustrate the process by which FRET is first identified and then quantified by our high-content screening system

M3 - Conference paper

SN - 0277-786X

T3 - PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)

SP - 85919

EP - 85919

BT - Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi

PB - SPIE - INT SOC OPTICAL ENGINEERING

CY - BELLINGHAM

T2 - Conference on Imaging, Manipulation and Analysis of Biomolecules, Cells, and Tissues VI

Y2 - 1 January 2008

ER -

Matthews DR, Ameer-Beg SM, Barber P, Pierce GP, Newman RG, Vojnovic B et al. A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919. In Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues Vi. BELLINGHAM: SPIE - INT SOC OPTICAL ENGINEERING. 2008. p. 85919 - 85919. (PROCEEDINGS OF THE SOCIETY OF PHOTO-OPTICAL INSTRUMENTATION ENGINEERS (SPIE)).

A high-content screening platform utilizing polarization anisotropy and FLIM microscopy - art. no. 685919

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