NOVY, Jindrich, Pavlina BECVAROVA, Jirina SKORPIKOVA, Vojtech MORNSTEIN and Roman JANISCH. Discrete Fourier transform-based analysis of HeLa cell microtubules after ultrasonic exposure. Microscopy Reasearch Technique. Wiley-Liss, 2005, vol. 68, No 1, p. 1-5. ISSN 1059-910X.
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Basic information
Original name Discrete Fourier transform-based analysis of HeLa cell microtubules after ultrasonic exposure
Name in Czech Diskrétní fourierovská analýza miktubul bunk HeLa po ultrazvukové expozici
Authors NOVY, Jindrich (203 Czech Republic), Pavlina BECVAROVA (203 Czech Republic), Jirina SKORPIKOVA (203 Czech Republic), Vojtech MORNSTEIN (203 Czech Republic, guarantor) and Roman JANISCH (203 Czech Republic).
Edition Microscopy Reasearch Technique, Wiley-Liss, 2005, 1059-910X.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10610 Biophysics
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 2.323
RIV identification code RIV/00216224:14110/05:00012839
Organization unit Faculty of Medicine
UT WoS 000232563100001
Keywords in English cytoskeleton; microtubules; immunofluorescence; FFT analysis; ultrasound; image analysis
Tags Cytoskeleton, FFT analysis, Image analysis, immunofluorescence, Microtubules, ultrasound
Changed by Changed by: prof. RNDr. Vojtěch Mornstein, CSc., učo 2001. Changed: 12/12/2005 10:12.
Abstract
Cytoskeletal structures can be affected by ultrasound. We exposed HeLa cells to continuous ultrasound (1 MHz, 1 and 2 W.cm-2, 10 min. at 37 C). The microtubules were detected by the monoclonal antibody TU-01/SwAM/FITC, observed in a fluorescence microscope and photographed digitally. The images were processed by FFT magic software. The structure analysis is based on frequency domain filtering using discrete Fourier transform. We tried to design filters to extract information describing best the structural changes. The properties of the filter can be enhanced by direction filtering, i.e., extraction of a symmetric angular segment in the frequency domain centred on a zero frequency. The final image is a normalised sum of inverse FFTs of such segmented spectra. The method yielded the ratio of the area of microtubules to the total cell area. Assuming that the image background intensity is constant, we can use thresholding to detect areas occupied by the cells. The information about the area of the microtubules is contained in a wide range of higher intensities. Therefore, we use a gamma correction. The area occupied by microtubules is then considered an area with intensities above the selected threshold. There were tested three different filters. The mathematical method chosen seems sensitive enough for quantitative assessment of changes of the microtubular network.
Abstract (in Czech)
Cytoskeletální struktury mohou být ovlivnovany ultrazvukem. Bunky HeLa jsme vystavili spojitému ultrazvuku (1 MHz, 1-2 W.cm-2, 10 min. pri 37 C). Mikrotubuly byly detegovány monoklonální protilátkou TU-01/SwAM/FITC, pozorovány ve fluorescencním mikroskopu a digitálne fotografovány. Obrázky byla zpracovány pomocí software "FTT Magic". Strukturní analýza je založena filtrování frekvencních domén pomocí diskrétní fourierovské transformace. Pokusili jsme se vytvorit filtr pro získání informace nejlépe popisující zmeny struktury. Vlastnosti filtru mohou být zesíleny smerovým filtrováním, t.j. vyjmutím symetrického úhlového segmentu ve frekvencní oblasti centrované na nulu. Výsledný obraz je normalizovaným souctem inverzních FTT takto segmentovaných spekter. Metoda poskytla pomer plochy mikrotubulu k celkové ploše bunky. Za predpokladu konstantní úrovne intenzity pozadí, použili jsme pro urení ploch bunek prahování. Informace o ploše mikrotubulu je obsdažena v širokém spektru vyšších intenzit. Proto bylo použito gama-korekce. Plocha považovaná za mikrotubuly pak byla považována za plochu s intenzitami nad zvoleným prahem. Byly testovány tri ruzné filtry. Zvolená matematická metoda se zdá být dostatecne citlivá pro kvantitativní hodnocení zmen síte mikrotubulu.
Links
GA301/01/1102, research and development projectName: Ovlivnění cytoskeletu normálních a nádorových buněk fyzikálními faktory a látkami s protinádorovou aktivitou
Investor: Czech Science Foundation, Influencing the cytoskeleton of normal and tumour cells by physical factors and substances with anti-tumour activity
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