Finding the bottom and using it: Offsets and sensitivity in the detection of low intensity values in vivo with 2-photon microscopy

dc.contributor.authorSandoval, Ruben M.
dc.contributor.authorWang, Exing
dc.contributor.authorMolitoris, Bruce A.
dc.contributor.departmentDepartment of Medicine, School of Medicineen_US
dc.date.accessioned2015-10-29T21:34:48Z
dc.date.available2015-10-29T21:34:48Z
dc.date.issued2014-03-01
dc.description.abstractMaximizing 2-photon parameters used in acquiring images for quantitative intravital microscopy, especially when high sensitivity is required, remains an open area of investigation. Here we present data on correctly setting the black level of the photomultiplier tube amplifier by adjusting the offset to allow for accurate quantitation of low intensity processes. When the black level is set too high some low intensity pixel values become zero and a nonlinear degradation in sensitivity occurs rendering otherwise quantifiable low intensity values virtually undetectable. Initial studies using a series of increasing offsets for a sequence of concentrations of fluorescent albumin in vitro revealed a loss of sensitivity for higher offsets at lower albumin concentrations. A similar decrease in sensitivity, and therefore the ability to correctly determine the glomerular permeability coefficient of albumin, occurred in vivo at higher offset. Finding the offset that yields accurate and linear data are essential for quantitative analysis when high sensitivity is required.en_US
dc.eprint.versionAuthor's manuscripten_US
dc.identifier.citationSandoval, R. M., Wang, E., & Molitoris, B. A. (2014). Finding the bottom and using it: Offsets and sensitivity in the detection of low intensity values in vivo with 2-photon microscopy. Intravital, 2(1), e23674. http://doi.org/10.4161/intv.23674en_US
dc.identifier.urihttps://hdl.handle.net/1805/7299
dc.publisherTaylor & Francis Onlineen_US
dc.relation.isversionof10.4161/intv.23674en_US
dc.relation.journalIntraVitalen_US
dc.rightsPublisher Policyen_US
dc.sourcePMCen_US
dc.subjectblack levelen_US
dc.subjectoffseten_US
dc.subjectsensitivityen_US
dc.subjectlow intensityen_US
dc.subject2-photonen_US
dc.subjectalbuminen_US
dc.subjectpermeabilityen_US
dc.titleFinding the bottom and using it: Offsets and sensitivity in the detection of low intensity values in vivo with 2-photon microscopyen_US
dc.typeArticleen_US
Files
Original bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
nihms606483.pdf
Size:
2.69 MB
Format:
Adobe Portable Document Format
Description:
Article
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.88 KB
Format:
Item-specific license agreed upon to submission
Description: