CONFOCAL MICROSCOPY

GROUP MEMBERS
SHAMNAD SHAMNA
NIZAR SUHAS
GLADIS BABY GLADIS
TARAK RAM
INTRODUCTION
 Confocal microscopy-most frequently confocal laser scanning
microscopy(CLSM)or laser confocal scanning
microscopy(LCSM).
 It is an optical imaging technique for increasing optical
resolution and contrast of a micrograph by means of using a
spatial pinhole to block out-of-focus light in image
formation.
PRINCIPLE OF CONFOCAL MICROSCOPE

 Florescence is the priniciple in confocal microscope. Florescence

is the phenomenon by which,when a molecule is shined with a
higher energy light, a light of lesser energy is emitted.
Working of confocal microscope
WORKING OF CONFOCAL MICROSCOPE
 Similar to the widefield microscope,the confocal microscope
uses fluorescence optics.
 Insead of ilumimating the whole sample at
 light is focused onto a defined spot at a specific depth within
the sample.
 Excitation and emission light pathways in a basic confocal
microscope configuration.
 A laser is used to provide the excitation light (in order to get
very high intensities). The laser light (blue) reflects off a
dichroic mirror. From there, the laser hits two mirrors which
are mounted on motors; these mirrors scan the laser across
the sample. Dye in the sample fluoresces, and the emitted
light (green) gets descanned by the same mirrors that are
used to scan the excitation light (blue) from the laser. The
emitted light passes through the dichroic and is focused onto
the pinhole. The light that passes through the pinhole is
measured by a detector, ie., a photomultiplier tube.