Appl Opt. 1999 Oct 1;38(28):5995-6003.

Imaging properties in two-photon excitation microscopy and effects of refractive-index mismatch in thick specimens.

de Grauw CJ, Vroom JM, van der Voort HT, Gerritsen HC.

Source

Molecular Biophysics, Utrecht University, P.O. Box 80.000, 3508 TA Utrecht, The Netherlands. k.degrauw@phys.uu.nl

Abstract

The detrimental effects of a refractive-index mismatch on the image formation in a two-photon microscope were investigated. Point-spread functions (PSF's) were recorded with an oil-immersion objective numerical aperture (NA) of 1.3 and a water-immersion objective NA of 1.2 in an aqueous sample at different depths. For the oil-immersion objective the enlargement of the PSF volume with increasing depth yields an axial and a lateral loss in resolution of approximately 380% and 160%, respectively, at a 90-microm depth in the sample. For the water-immersion objective no resolution decrease was found. Measurements on a thick aqueous biofilm sample shows the importance of matching the refractive index between immersion fluid and sample. With a good match, no loss in image resolution is observed.

PMID:: 18324119; [PubMed]

LinkOut - more resources

Full Text Sources

Supplemental Content

Save items

Related citations in PubMed

Imaging of optically thick specimen using two-photon excitation microscopy. [Microsc Res Tech. 1999]
Imaging of optically thick specimen using two-photon excitation microscopy.
Gerritsen HC, De Grauw CJ. Microsc Res Tech. 1999 Nov 1; 47(3):206-9.
Performances of high numerical aperture water and oil immersion objective in deep-tissue, multi-photon microscopic imaging of excised human skin. [Microsc Res Tech. 2004]
Performances of high numerical aperture water and oil immersion objective in deep-tissue, multi-photon microscopic imaging of excised human skin.
Dong CY, Yu B, Kaplan PD, So PT. Microsc Res Tech. 2004 Jan 1; 63(1):81-6.
A new high-aperture glycerol immersion objective lens and its application to 3D-fluorescence microscopy. [J Microsc. 2002]
A new high-aperture glycerol immersion objective lens and its application to 3D-fluorescence microscopy.
Martini N, Bewersdorf J, Hell SW. J Microsc. 2002 May; 206(Pt 2):146-51.
Effects of different immersion media in multiphoton imaging of the epithelium and dermis of human skin. [Microsc Res Tech. 2006]
Effects of different immersion media in multiphoton imaging of the epithelium and dermis of human skin.
Hsiao CY, Sun Y, Chen WL, Tung CK, Lo W, Su JW, Lin SJ, Jee SH, Jan GJ, Dong CY. Microsc Res Tech. 2006 Dec; 69(12):992-7.
A procedure to determine the correct thickness of an object with confocal microscopy in case of refractive index mismatch. [J Microsc. 2005]
A procedure to determine the correct thickness of an object with confocal microscopy in case of refractive index mismatch.
Kuypers LC, Decraemer WF, Dirckx JJ, Timmermans JP. J Microsc. 2005 Apr; 218(Pt 1):68-78.

See reviews... See all...

Cited by 3 PubMed Central articles

Integrated holographic system for all-optical manipulation of developing embryos. [Biomed Opt Express. 2011]
Integrated holographic system for all-optical manipulation of developing embryos.
Torres-Mapa ML, Antkowiak M, Cizmarova H, Ferrier DE, Dholakia K, Gunn-Moore FJ. Biomed Opt Express. 2011 Jun 1; 2(6):1564-75. Epub 2011 May 16.
Maximum imaging depth of two-photon autofluorescence microscopy in epithelial tissues. [J Biomed Opt. 2011]
Maximum imaging depth of two-photon autofluorescence microscopy in epithelial tissues.
Durr NJ, Weisspfennig CT, Holfeld BA, Ben-Yakar A. J Biomed Opt. 2011 Feb; 16(2):026008.
Spectrally resolved multiphoton imaging of in vivo and excised mouse skin tissues. [Biophys J. 2007]
Spectrally resolved multiphoton imaging of in vivo and excised mouse skin tissues.
Palero JA, de Bruijn HS, van der Ploeg van den Heuvel A, Sterenborg HJ, Gerritsen HC. Biophys J. 2007 Aug 1; 93(3):992-1007. Epub 2007 Apr 20.

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

Imaging properties in two-photon excitation microscopy and effects of refractive...
Imaging properties in two-photon excitation microscopy and effects of refractive-index mismatch in thick specimens.
Appl Opt. 1999 Oct 1 ;38(28):5995-6003.

PubMed
Advances in the molecular biology of malignant mesothelioma.
Advances in the molecular biology of malignant mesothelioma.
Acta Med Okayama. 2008 Feb ;62(1):1-7.

PubMed
Generation of monoclonal antibodies against soluble human T cell receptor polype...
Generation of monoclonal antibodies against soluble human T cell receptor polypeptides.
Eur J Immunol. 1991 Sep ;21(9):2111-9.

PubMed
Targeting melanocortin receptors: an approach to treat weight disorders and sexu...
Targeting melanocortin receptors: an approach to treat weight disorders and sexual dysfunction.
Nat Rev Drug Discov. 2008 Apr ;7(4):307-23.

PubMed
Quantitative proteomics using stable isotope labeling with amino acids in cell c...
Quantitative proteomics using stable isotope labeling with amino acids in cell culture.
Nat Protoc. 2008 ;3(3):505-16.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to:

Imaging properties in two-photon excitation microscopy and effects of refractive-index mismatch in thick specimens.

Source

Abstract

LinkOut - more resources

Full Text Sources

Supplemental Content

Save items

Related citations in PubMed

Cited by 3 PubMed Central articles

Related information

Recent activity

Simple NCBI Directory

Getting Started

Resources

Popular

Featured

NCBI Information