Confocal Microscopy Principles

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Nipkow disk confocal microscope facilitate Ca 2+ waves in cardiomyocytes loaded with fluo-3 Dr. Marisa Jaconi Geneva studies of fast processes Image capture at 33 Hz using an intensified camera (Coolsnap Cascade from Photometrics)
Other beam scanning techniques Slit scanning : a new approach in confocal microscopy •The circular laser beam is transformed to a line which scan the sample in only one direction •The emitted fluorescence of that line passed through a confocal line pinhole •This line (512 pixels) is detected by a ultrafast line CCD detector Scan speeds of 100 frames/s can be achieved
Page 1 and 2: Confocal Microscopy Dr. Serge Arnau
Page 3 and 4: Light source in focus Sample (objec
Page 5 and 6: Light source Confocal microscope pr
Page 7 and 8: How does a fluorescence microscope
Page 9 and 10: Advantage of fluorescence confocal
Page 11 and 12: LASER Light Amplification by Stimul
Page 13 and 14: Beam scanning Majority of laser sca
Page 15 and 16: Photomultiplier Tubes (PMT) The spe
Page 17 and 18: Scans averaging reduces noise Avera
Page 19 and 20: Airy disk and Resolution The radius
Page 21 and 22: Pinhole and Resolution Confocal pin
Page 23 and 24: Pinhole and Resolution Confocal PSF
Page 25 and 26: Z Optical sectionning Y X 5 4 3 2 1
Page 27 and 28: Sampling in confocal microscopy 260
Page 29 and 30: Sampling in confocal microscopy The
Page 31 and 32: Ideal emission separation Red emiss
Page 33 and 34: Crosstalk problems To avoid bleed-t
Page 35 and 36: Spectral separation Fluorescence se
Page 37 and 38: FRAP experiments FRAP-recording for
Page 39 and 40: Other beam scanning techniques Mult
Page 41: Nipkow disk confocal microscope fac

Confocal Microscopy Principles

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