Single-Cell Fluorescence Spectroscopy


T. Levitner


Institute of Physical Biology, University of South Bohemia,

 Zámek 136,CZ-373 33 Nové Hrady


We are developing software for spectrum analysis measured in single living cell. We want to automatize analysis of measured data and to accelerate generating of results.


We measure cells of trichodesmia. We are measuring changes in spectra of one cell selected in the sample. Cell is irradiated by the blue light and intensity is changed according to the protocol. Spectrum is measured by spectrometer in range from 177 [nm] to 992 [nm] (from IR to UV). Output of measurement is the large table with light intensities measured on different wavelengths (rows) and time (columns) of measurement. Fluorescent parameters like FM, F0, FS are calculated of this table. Due the size of this table cannot be used common available tools like MS Excel or OpenOffice Calc etc.

Therefore I am developing program in the Matlab environment, which allows to process this large data set and to automatize generating of desired results. Program was named ‘Sandra’.

Illustrative picture displays table in 3D representation. This table has size 1044x1630 values. This is repeated measurement several times during the day. Manual processing of one table using MS Excel takes one day in the case that we use only of part measured spectra and required spectra is averaged. We are able to do the same and without restriction which we mentioned in several minutes using our software Sandra.


Up to now we have solved head parts like reading data of file, visualization data in 3D and partially plotting of fluorescent parameters in depend on wavelength.


The aim of those calculations is to express fluorescent parameters like functions wavelength (FM(λ), F0(λ),FS(λ),...) of measured data. Later we will concentrate on calculation time delays between excitation light and emission light of cell, which will allow us to better understand to elementary processes in metabolism of living cell.