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Title: Two-photon excited UV fluorescence for protein crystal detection

Abstract

Complementary measurements using SONICC and TPE-UVF allow the sensitive and selective detection of protein crystals. Two-photon excited ultraviolet fluorescence (TPE-UVF) microscopy is explored for sensitive protein-crystal detection as a complement to second-order nonlinear optical imaging of chiral crystals (SONICC). Like conventional ultraviolet fluorescence (UVF), TPE-UVF generates image contrast based on the intrinsic fluorescence of aromatic residues, generally producing higher fluorescence emission within crystals than the mother liquor by nature of the higher local protein concentration. However, TPE-UVF has several advantages over conventional UVF, including (i) insensitivity to optical scattering, allowing imaging in turbid matrices, (ii) direct compatibility with conventional optical plates and windows by using visible light for excitation, (iii) elimination of potentially damaging out-of-plane UV excitation, (iv) improved signal to noise through background reduction from out-of-plane excitation and (v) relatively simple integration into instrumentation developed for SONICC.

Authors:
; ;  [1]
  1. Purdue University, 560 Oval Drive, West Lafayette, IN 47907 (United States)
Publication Date:
OSTI Identifier:
22351245
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section D: Biological Crystallography
Additional Journal Information:
Journal Volume: 67; Journal Issue: Pt 10; Other Information: PMCID: PMC3176620; PMID: 21931215; PUBLISHER-ID: mh5044; OAI: oai:pubmedcentral.nih.gov:3176620; Copyright (c) International Union of Crystallography 2011; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0907-4449
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; COMPATIBILITY; CRYSTALS; EXCITATION; FLUORESCENCE; HARMONIC GENERATION; IMAGES; MICROSCOPY; NOISE; PHOTONS; PLATES; REDUCTION; SCATTERING; SIGNALS; VISIBLE RADIATION

Citation Formats

Madden, Jeremy T., DeWalt, Emma L., and Simpson, Garth J., E-mail: gsimpson@purdue.edu. Two-photon excited UV fluorescence for protein crystal detection. Denmark: N. p., 2011. Web. doi:10.1107/S0907444911028253.
Madden, Jeremy T., DeWalt, Emma L., & Simpson, Garth J., E-mail: gsimpson@purdue.edu. Two-photon excited UV fluorescence for protein crystal detection. Denmark. doi:10.1107/S0907444911028253.
Madden, Jeremy T., DeWalt, Emma L., and Simpson, Garth J., E-mail: gsimpson@purdue.edu. Sat . "Two-photon excited UV fluorescence for protein crystal detection". Denmark. doi:10.1107/S0907444911028253.
@article{osti_22351245,
title = {Two-photon excited UV fluorescence for protein crystal detection},
author = {Madden, Jeremy T. and DeWalt, Emma L. and Simpson, Garth J., E-mail: gsimpson@purdue.edu},
abstractNote = {Complementary measurements using SONICC and TPE-UVF allow the sensitive and selective detection of protein crystals. Two-photon excited ultraviolet fluorescence (TPE-UVF) microscopy is explored for sensitive protein-crystal detection as a complement to second-order nonlinear optical imaging of chiral crystals (SONICC). Like conventional ultraviolet fluorescence (UVF), TPE-UVF generates image contrast based on the intrinsic fluorescence of aromatic residues, generally producing higher fluorescence emission within crystals than the mother liquor by nature of the higher local protein concentration. However, TPE-UVF has several advantages over conventional UVF, including (i) insensitivity to optical scattering, allowing imaging in turbid matrices, (ii) direct compatibility with conventional optical plates and windows by using visible light for excitation, (iii) elimination of potentially damaging out-of-plane UV excitation, (iv) improved signal to noise through background reduction from out-of-plane excitation and (v) relatively simple integration into instrumentation developed for SONICC.},
doi = {10.1107/S0907444911028253},
journal = {Acta Crystallographica. Section D: Biological Crystallography},
issn = {0907-4449},
number = Pt 10,
volume = 67,
place = {Denmark},
year = {2011},
month = {10}
}