摘要:
Fluorescence generated by multiphoton excitation can be observed simultaneously using multiple beams, and multiple points can be observed simultaneously with a high signal-to-noise ratio with low invasiveness. Provided is a laser microscope 1 including: modulation units 5 and 6 that apply different modulations to a plurality of ultrashort-pulse laser light beams of the same type emitted from a light source unit 3; an illumination optical system 7 that simultaneously focuses the plurality of ultrashort-pulse laser light beams, to which the different modulations are applied by the modulation units 5 and 6, onto different positions of a sample O; a fluorescence detecting device 8 that detects fluorescence generated at a focal position of each ultrashort-pulse laser light beam and performs photoelectric conversion of the fluorescence; and a demodulation unit 9 that demodulates an output from the fluorescence detecting device 8 based on modulation information from the modulation units 5 and 6.
摘要:
An apparatus (10) for structured illumination microscopy (SIM) comprises a pulsed femtosecond MiTai laser (11) operable to generate a pulsed beam. The beam pulses are directed on to a specimen (12) via an optical arrangement including a beam steering apparatus comprising a pair of acousto-optic deflectors AODx, AODy, each operable to vary the deflection angle of the beam in response to variation in the frequency of an applied acoustic deflection signal,and an acousto-optic modulator AOM. The frequency of the acoustic deflection signals applied to the AODs and/or the frequency of the acoustic compensation signal applied to the AOM in the present invention is dynamically modulated. This dynamic modulation can increase the effective AODxy scan angle in each direction by about 4 mrads (equivalent to a 15% increase in the area of the field of view). Furthermore, dynamic modulation of the compensation frequency also improves the evenness of the illumination over the field of view.
摘要:
Die Erfindung betrifft eine Auswerteschaltung (1) zum Auswerten der elektrischen Signale (A) eines zur Detektion von Photonen ausgebildeten optoelektronischen Detektors (32) bei der Aufnahme von Fluoreszenzereignissen unter Anregung mittels Lichtpulsen, umfassend einen Analog-Digital-Wandler (2) oder einen Trigger mit einem Eingang zur Verbindung mit einem Ausgang des Detektors (32) und mit einem Ausgang zur Ausgabe eines Digitalwerts (a), ein Schieberegister (3), wobei die Auswerteschaltung (1) so konfiguriert ist, dass während eines Beleuchtungszyklus zu unterschiedlichen Zeitpunkten aufgenommene Lichtintensitäten unabhängig voneinander in den Stufen des Schieberegister gespeichert werden. Die Erfindung betrifft ferner die Verwendung der Auswerteschaltung (1) zur zeitlichen Trennung von Fluoreszenz- und Streulicht von Fluoreszenzereignissen sowie ein Verfahren zum Aufzeichnen von Fluoreszenzereignissen.
摘要:
The invention relates to a laser scanning microscope (10) having: focusing means (15) having a focal plane (29) and comprising at least one optical element for focusing a laser beam (13); drive means (18) for displacing the at least one optical element of the focusing means (15) for changing the position of the focal plane (29), and deflecting means (14) for deflecting the laser beam (13). The microscope comprises a control system (32) configured to carry out the steps of: providing a periodical drive signal for the drive means (18); obtaining time dependant displacement data of the at least one optical element of the focusing means (15) in response to the periodical drive signal of the drive means (18); providing a response function (z(t)) using the time dependant displacement data, calculating a drive signal for the deflecting means (14) using the response function (z(t)) to move the focal volume (30) of the laser beam (13) along a given 3D trajectory (48) within a sample to be examined. The invention further relates to a method for carrying out such a scanning operation along a 3D trajectory (48). ˙