Phase-contrast microscopy, technique

At this point it is worth comparing the different techniques of contrast enliancements discussed so far. They represent spatial filtering teclmiques which mostly affect the zeroth order dark field microscopy, which eliminates the zeroth order, the Schlieren method (not discussed here), which suppresses the zerotii order and one side band and, finally, phase contrast microscopy, where the phase of the zeroth order is shifted by nil and its intensity is attenuated. 

The direct visnalization of microstructure may be accomplished by various forms of microscopy. Recent refinements in microscopy techniques are epitomized by video-enhanced interference phase-contrast microscopy, which is emerging as a workhorse probe for colloidal suspensions and other microstructnred liqnids. 

Reflected light Microscopy can be used for examining the texture of solid opaque polymers. Materials which can be prepared as thin films are generally examined by transmitted light. Two common techniques used are (i) polarised-light Microscopy, and (ii) phase contrast Microscopy. 

Later, differential interference microscopy was developed, enabling the detection of difference in levels as sensitively as phase contrast microscopy, and, because this technique was easier to use, it came to be used in preference to the former techniques [6]. Differential interference microscopy is superior to phase contrast microscopy in the observation of vicinal or curved surfaces, which are impossible to observe under a phase contrast microscope because the contrast is too high. 

Powerful methods that have been developed more recently, and are currently used to observe surface micro topographs of crystal faces, include scanning tunnel microscopy (STM), atomic force microscopy (AFM), and phase shifting microscopy (PSM). Both STM and AFM use microscopes that (i) are able to detect and measure the differences in levels of nanometer order (ii) can increase two-dimensional magnification, and (iii) will increase the detection of the horizontal limit beyond that achievable with phase contrast or differential interference contrast microscopy. The presence of two-dimensional nuclei on terraced surfaces between steps, which were not observable under optical microscopes, has been successfully detected by these methods [8], [9]. In situ observation of the movement of steps of nanometer order in height is also made possible by these techniques. However, it is possible to observe step movement in situ, and to measure the surface driving force using optical microscopy. The latter measurement is not possible by STM and AFM. 

Air drawn through a 0.8 to 1.2 mm cellulose ester membrane filter, asbestos fibers counted by positive phase contrast microscopy technique sample prepared by acetone/triacetin method (NIOSH Method 7400, 1985). 

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