Analizatory DNA/PCR typu lab-chip z detekcją fluorymetryczną

• Autorzy: Walczak R.

Warianty tytułu

EN

Lab-on-a-chip DNA/PCR analyzers with fluorometric detection

• Języki publikacji: PL

Abstrakty

PL

W artykule przedstawiono przegląd analizatorów DNA wykorzystujących technikę real-time PCR i lab-chipy. Zaprezentowano opis opracowanej metodologii i instrumentarium do detekcji fluorescencji z wykorzystaniem miniaturowych komponentów optoelektronicznych i "inteligentnego" oprogramowania analizującego. Przedstawiono dwa przykłady miniaturowych analizatorów real-time PCR/DNA opracowanych w ramach projektów europejskich i krajowych wykorzystujących lab-chipy i nowatorską metodę detekcji fluorymetrycznej.

EN

The paper presents a novel miniaturized optical instrumentation for fluorescence excitation and detection for miniaturized real-time PCR analyzers using lab-on-a-chip (LOC) devices. Application of miniaturized semiconductor laser to fluorescence excitation, CCD-minicamera as fluorescence photodetector and specialized software for optical signal conditioning led to development of low-cost and highly sensitive optical instrumentation. To carry out full characterization of the optical instrumentation, miniaturized thermocycler co-working with LOC was built. The optical instrumentation was tested with three LOC made of different materials and technologies, giving proper detection of fluorescence signals during real-time PCR. Finally, two miniaturized devices for real-time PCR detection and identification of DNA and utilizing described here novel optical instrumentation were briefly described. The scheme and technical realization of the novel instrumentation in laboratory version is shown in Fig. 4. The detection limit of DNA was about 0,1 ng/ml (Fig. 5). It was also found that the optical instrumentation co-works with different constructions of lab-on-a-chips for DNA real-time PCR amplification (Fig. 6). The developed optical instrumentation was successfully applied to two miniaturized devices for DNA detection and identification by real-time PCR. Technical realizations of the devices and views of the lab-on-a-chips are shown in Figs. 7 and 9. In both cases, proper detection of fluorescence signals generated during real-time PCR of Campylobacter j. DNA (Fig. 8) or complementary DNA (Fig. 10) were observed. It confirmed high sensitivity of the developed optical instrumentation.

Słowa kluczowe

PL

real-time PCR; lab-chip; fluorescencja

EN

real-time PCR; lab-chip; fluorescence

• Wydawca: Wydawnictwo PAK
• Czasopismo: Pomiary Automatyka Kontrola
• Rocznik: 2010
• Tom: R. 56, nr 7
• Strony: 805--808
• Opis fizyczny: Bibliogr. 14 poz., rys., wykr.

Twórcy

autor

Walczak R.

• Politechnika Wrocławska, Wydział Elektroniki Mikrosystemów i Fotoniki, Zakład Mikroinżynierii i Fotowoltaiki, ul. Janiszewskiego 11/16, 50-372 Wrocław,

Bibliografia

• [1] Advan. Physiol. Educ., American Physiological Society, 29, 2005.
• [2] Biomedical Microdevices, Springer, 7:4, 2005.
• [3] Nucleid Acid Research, Oxford Journals, vol. 34, no 11, 2006.
• [4] Sensors and Actuators B, Elsevier, 107, 2005.
• [5] Biomedical Microdevices, Springer, 9, 2007.
• [6] Sensors and Actuators B, Elsevier, 100, 2004.
• [7] Proc. of μTAS 2006, Transducers Research Foundation, Tokyo, Japonia.
• [8] Biosensors and Bioelectronics 21, 2006.
• [9] Proc. of MEMS 2006, Transducers Research Foundation, Istambuł, Turcja.
• [10] Lab Chip, RSC Publishing, 5, 2005.
• [11] Proc. of μTAS 2007, Transducers Research Foundation Paryż, Francja.
• [12] Proc. of μTAS 2008, Transducers Research Foundation, San Diego, USA.
• [13] Lab Chip, RSC Publishing , vol. 9, iss. 11, 2009.
• [14] Proc. of μTAS 2009, Transducers Research Foundation, South Korea.