Detailed Information on Publication Record
2007
Optimizing light throughput in image cytometry
KOZUBEK, Michal and Jakub KULAVIAKBasic information
Original name
Optimizing light throughput in image cytometry
Name in Czech
Optimalizace světelné propustnosti v obrazové cytometrii
Authors
Edition
Analytical Cytometry IV: International Conference, 2007
Other information
Language
English
Type of outcome
Vyžádané přednášky
Field of Study
20200 2.2 Electrical engineering, Electronic engineering, Information engineering
Country of publisher
Czech Republic
Confidentiality degree
není předmětem státního či obchodního tajemství
Organization unit
Faculty of Informatics
Keywords in English
fluorescence microscopy; light throughput
Tags
International impact
Změněno: 8/1/2009 19:39, prof. RNDr. Michal Kozubek, Ph.D.
V originále
Image cytometry applications often suffer from the lack of light due to the use of fluorescence mode. Low light levels typically cause poor signal-to-noise ratio or long exposure times. Signal level in fluorescence set-up is determined by many factors, such as power of the light source and its spectral characteristics, transmission curves of excitation/emission filters, transmission curves of and dichroic/polychroic mirrors, excitation/emission spectra of imaged fluorochrome(s), numerical aperture of the objective and its transmission curve, quantum efficiency curve of the light detector. Thus, resulting light throughput is given by the composition of spectral characteristics (functions of wavelength) of many components. Proper computation and optimization of light throughput is a non-trivial task that requires knowledge of all these curves for all available hardware components as well as all available fluorochromes. Such computation is unfortunately not performed by the users and is typically replaced, for example, by visual comparison of fluorochrome spectral curves with spectral curves of optical filters when choosing fluorochromes or filters. Quantum efficiency curve of the detector or spectral characteristics of the white light source are usually not taken into account at all. In our laboratory, we have developed software for the computation of the overall light throughput of an optical set-up. It can assist users in choosing hardware components for their system as well as choosing suitable fluorochromes. For each component the user enters the available choices (e.g. list of available optical filters, list of available fluorochromes, list of available cameras in camera-based systems, etc.) and the computer suggests a list of suitable combinations ordered according to the resulting signal level. Contrast between signal from emitted fluorescence and signal from remaining excitation light is also taken into account. The software runs on a web server and can be used from a web browser for: choosing a suitable fluorochrome when ordering probes or staining kits, choosing suitable optical filters (excitation/emission) to be installed or purchased, choosing suitable mirrors (dichroic/polychroic) to be installed or purchased, choosing suitable light detector (especially in camera-based systems), comparing light throughput and contrast for fluorochromes imaged simultaneously, comparing light throughput and contrast for different combinations of hardware components available in the lab. This research has been supported by the Ministry of Education of the Czech Republic (Projects No. MSM0021622419 and No. LC535).
In Czech
Přednáška se zabývá optimalizací světelné propustnosti v obrazové cytometrii
Links
| LC535, research and development project |
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| MSM0021622419, plan (intention) |
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