Confocal Microscopy Principles

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Scanning speed influences image quality pixel dwell time 3.2 µs pixel dwell time 25.6 µs Better signal to noise ratio with low scanning speeds but samples are more exposed to the laser beam 2 µm Muntjac cells – Alexa 555 anti OX Phos complex V inh prot
Scans averaging reduces noise Average of 2 scans Average of 8 scans But greatly reduce the frame rate 10 µm Muntjac cells – Alexa 488 phalloidin
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: Photomultiplier Tubes (PMT) The spe
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 and 42: Nipkow disk confocal microscope fac
Page 43 and 44: Other beam scanning techniques Slit

Confocal Microscopy Principles

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