Tirf single molecule analysis and method of sequencing...

G - Physics – 01 – N

Patent

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G01N 21/64 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2588122

An apparatus for analyzing the presence of a single molecule comprises a sample plate (30~) having a sample located thereon. In one embodiment the apparatus comprises two lasers (46~, 46~~) for providing illumination light at two different wavelengths suitable for exciting fluorescence in the sample, a collimator comprising a band-pass filter (50~, 55~~), a diverging lens (54~, 54~~), a field stop (62~), and a converging lens (66~) for directing the illunination light through a total internal reflection objective (74~) onto the sample, and means (34~, 34~~) for detecting a fluorescent image produced by said sample in response to the illunination light. In one embodiment the apparatus further comprises an auto-focus module comprising a laser (38~), a beam splitter cube (86~), a converging lens (90~), and an auto-focus dichroec (94~) for maintaining focus of the objective on the sample.

Dans un aspect, l'invention concerne un appareil destiné à analyser la présence de molécule unique à l'aide d'une réflexion interne totale (TIRF). Dans un mode de réalisation, un appareil pour une analyse de molécule unique comprend un support présentant un échantillon situé sur celui-ci ; deux sources lumineuses à des longueurs d'ondes distinctes, un collimateur destiné à diriger la lumière sur l'échantillon à travers un objectif à réflexion interne totale ; un récepteur destiné à recevoir une émission fluorescente produite par une molécule unique dans l'échantillon en réponse à la lumière ; et un détecteur destiné à détecter chacune des longueurs d'ondes dans l'émission fluorescente. Dans un autre mode de réalisation, l'appareil comprend en outre un laser de focalisation destiné à maintenir le foyer de l'objectif sur l'échantillon.

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