Claudio Vinegoni, Ph.D. | Born normalization for fluorescence optical projection tomography for whole heart imaging

J Vis Exp
Born normalization for fluorescence optical projection tomography for whole heart imaging

Vinegoni^#, C., Razansky, D., Figueiredo, J. L., Fexon, L., Pivoravov, M., Nahrendorf, M., Ntziachristos, V., and Weissleder, R.

Journal of Visualized Experiments 2009

ABS PMID DOI PMCID CITE HTML PDF

Optical projection tomography is a three-dimensional imaging technique that has been recently introduced as an imaging tool primarily in developmental biology and gene expression studies. The technique renders biological sample optically transparent by first dehydrating them and then placing in a mixture of benzyl alcohol and benzyl benzoate in a 2:1 ratio (BABB or Murray s Clear solution). The technique renders biological samples optically transparent by first dehydrating them in graded ethanol solutions then placing them in a mixture of benzyl alcohol and benzyl benzoate in a 2:1 ratio (BABB or Murray s Clear solution) to clear. After the clearing process the scattering contribution in the sample can be greatly reduced and made almost negligible while the absorption contribution cannot be eliminated completely. When trying to reconstruct the fluorescence distribution within the sample under investigation, this contribution affects the reconstructions and leads, inevitably, to image artifacts and quantification errors.. While absorption could be reduced further with a permanence of weeks or months in the clearing media, this will lead to progressive loss of fluorescence and to an unrealistically long sample processing time. This is true when reconstructing both exogenous contrast agents (molecular contrast agents) as well as endogenous contrast (e.g. reconstructions of genetically expressed fluorescent proteins).
@article{2009-JOVE-1, author= {Vinegoni<sup>#</sup>, C. and Razansky, D. and Figueiredo, J. L. and Fexon, L. and Pivoravov, M. and Nahrendorf, M. and Ntziachristos, V. and Weissleder, R.}, title= {Born normalization for fluorescence optical projection tomography for whole heart imaging}, journal= {Journal of Visualized Experiments}, alternatejournal = {J Vis Exp}, year = {2009}, volume = {2}, number = {28}, pages = {1389}, doi = {10.3791/1389}, pmcid = {PMC2794886}, pmid= {19578329} }
19578329

PMC2794886

10.3791/1389

Abstract

"Optical projection tomography is a three-dimensional imaging technique that has been recently introduced as an imaging tool primarily in developmental biology and gene expression studies. The technique renders biological sample optically transparent by first dehydrating them and then placing in a mixture of benzyl alcohol and benzyl benzoate in a 2:1 ratio (BABB or Murray s Clear solution). The technique renders biological samples optically transparent by first dehydrating them in graded ethanol solutions then placing them in a mixture of benzyl alcohol and benzyl benzoate in a 2:1 ratio (BABB or Murray s Clear solution) to clear. After the clearing process the scattering contribution in the sample can be greatly reduced and made almost negligible while the absorption contribution cannot be eliminated completely. When trying to reconstruct the fluorescence distribution within the sample under investigation, this contribution affects the reconstructions and leads, inevitably, to image artifacts and quantification errors.. While absorption could be reduced further with a permanence of weeks or months in the clearing media, this will lead to progressive loss of fluorescence and to an unrealistically long sample processing time. This is true when reconstructing both exogenous contrast agents (molecular contrast agents) as well as endogenous contrast (e.g. reconstructions of genetically expressed fluorescent proteins)."

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For attribution in academic contexts, please cite this work as:

Vinegoni^#, C., Razansky, D., Figueiredo, J. L., Fexon, L., Pivoravov, M., Nahrendorf, M., Ntziachristos, V., & Weissleder, R. (2009). Born normalization for fluorescence optical projection tomography for whole heart imaging. Journal of Visualized Experiments, 2(28), 1389. https://doi.org/10.3791/1389