The Effects of Refractive Index Heterogeneity Within Kidney Tissue on Multiphoton Fluorescence Excitation Microscopy

Overview

Journal J Microsc

Specialty Radiology

Date 2010 Dec 2

PMID 21118239

Citations 8

Authors

P A Young

S G Clendenon

J M Byars

K W Dunn

Affiliations

Soon will be listed here.

Abstract

Although multiphoton fluorescence excitation microscopy has improved the depth at which useful fluorescence images can be collected in biological tissues, the reach of multiphoton fluorescence excitation microscopy is nonetheless limited by tissue scattering and spherical aberration. Scattering can be reduced in fixed samples by mounting in a medium whose refractive index closely matches that of the fixed material. Using optical 'clearing', the effects of refractive index heterogeneity on signal attenuation with depth are investigated. Quantitative measurements show that by mounting kidney tissue in a high refractive index medium, less than 50% of signal attenuates in 100 μm of depth.

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References

Zipfel W, Williams R, Webb W . Nonlinear magic: multiphoton microscopy in the biosciences. Nat Biotechnol. 2003; 21(11):1369-77. DOI: 10.1038/nbt899. View

Dong C, Koenig K, So P . Characterizing point spread functions of two-photon fluorescence microscopy in turbid medium. J Biomed Opt. 2003; 8(3):450-9. DOI: 10.1117/1.1578644. View

Oldham M, Sakhalkar H, Oliver T, Johnson G, Dewhirst M . Optical clearing of unsectioned specimens for three-dimensional imaging via optical transmission and emission tomography. J Biomed Opt. 2008; 13(2):021113. PMC: 2746042. DOI: 10.1117/1.2907968. View

Centonze V, White J . Multiphoton excitation provides optical sections from deeper within scattering specimens than confocal imaging. Biophys J. 1998; 75(4):2015-24. PMC: 1299873. DOI: 10.1016/S0006-3495(98)77643-X. View

Helmchen F, Denk W . Deep tissue two-photon microscopy. Nat Methods. 2005; 2(12):932-40. DOI: 10.1038/nmeth818. View

Pawley J . Limitations on optical sectioning in live-cell confocal microscopy. Scanning. 2002; 24(5):241-6. DOI: 10.1002/sca.4950240504. View

Oheim M, Beaurepaire E, Chaigneau E, Mertz J, Charpak S . Two-photon microscopy in brain tissue: parameters influencing the imaging depth. J Neurosci Methods. 2001; 111(1):29-37. DOI: 10.1016/s0165-0270(01)00438-1. View

Dunn A, Wallace V, Coleno M, Berns M, Tromberg B . Influence of optical properties on two-photon fluorescence imaging in turbid samples. Appl Opt. 2008; 39(7):1194-201. DOI: 10.1364/ao.39.001194. View

Clendenon S, Young P, Ferkowicz M, Phillips C, Dunn K . Deep tissue fluorescent imaging in scattering specimens using confocal microscopy. Microsc Microanal. 2011; 17(4):614-7. PMC: 4428593. DOI: 10.1017/S1431927611000535. View

10.

Beek J, Blokland P, Posthumus P, Aalders M, Pickering J, Sterenborg H . In vitro double-integrating-sphere optical properties of tissues between 630 and 1064 nm. Phys Med Biol. 1997; 42(11):2255-61. DOI: 10.1088/0031-9155/42/11/017. View

11.

Schilders , Gu . Limiting Factors on Image Quality in Imaging through Turbid Media under Single-photon and Two-photon Excitation. Microsc Microanal. 2000; 6(2):156-160. DOI: 10.1007/s100059910018. View

12.

Konig K . Multiphoton microscopy in life sciences. J Microsc. 2000; 200(Pt 2):83-104. DOI: 10.1046/j.1365-2818.2000.00738.x. View

13.

Millon S, Roldan-Perez K, Riching K, Palmer G, Ramanujam N . Effect of optical clearing agents on the in vivo optical properties of squamous epithelial tissue. Lasers Surg Med. 2006; 38(10):920-7. DOI: 10.1002/lsm.20451. View

14.

Doornbos R, Lang R, Aalders M, Cross F, Sterenborg H . The determination of in vivo human tissue optical properties and absolute chromophore concentrations using spatially resolved steady-state diffuse reflectance spectroscopy. Phys Med Biol. 1999; 44(4):967-81. DOI: 10.1088/0031-9155/44/4/012. View

15.

Plotnikov S, Juneja V, Isaacson A, Mohler W, Campagnola P . Optical clearing for improved contrast in second harmonic generation imaging of skeletal muscle. Biophys J. 2005; 90(1):328-39. PMC: 1367031. DOI: 10.1529/biophysj.105.066944. View

16.

Theer P, Denk W . On the fundamental imaging-depth limit in two-photon microscopy. J Opt Soc Am A Opt Image Sci Vis. 2006; 23(12):3139-49. DOI: 10.1364/josaa.23.003139. View

17.

Young P, Clendenon S, Byars J, Decca R, Dunn K . The effects of spherical aberration on multiphoton fluorescence excitation microscopy. J Microsc. 2010; 242(2):157-65. PMC: 4449278. DOI: 10.1111/j.1365-2818.2010.03449.x. View

18.

Cuccia D, Bevilacqua F, Durkin A, Merritt S, Tromberg B, Gulsen G . In vivo quantification of optical contrast agent dynamics in rat tumors by use of diffuse optical spectroscopy with magnetic resonance imaging coregistration. Appl Opt. 2003; 42(16):2940-50. DOI: 10.1364/ao.42.002940. View

19.

Cicchi R, Sampson D, Massi D, Pavone F . Contrast and depth enhancement in two-photon microscopy of human skin ex vivo by use of optical clearing agents. Opt Express. 2009; 13(7):2337-44. DOI: 10.1364/opex.13.002337. View

20.

Ying J, Liu F, Alfano R . Spatial distribution of two-photon-excited fluorescence in scattering media. Appl Opt. 2008; 38(1):224-9. DOI: 10.1364/ao.38.000224. View