Abstract
Optical microscopy promises researchers to see most tiny substances directly. However, the resolution of conventional microscopy is restricted by the diffraction limit. This makes it a challenge to observe subcellular processes happened in nanoscale. The development of super-resolution microscopy provides a solution to this challenge. Here, we briefly review several commonly used super-resolution techniques, explicating their basic principles and applications in biological science, especially in neuroscience. In addition, characteristics and limitations of each technique are compared to provide a guidance for biologists to choose the most suitable tool.
Highlights
Optical microscopy is one of the key, versatile and powerful tool in scientic researches, especially in biology
When light passes through an objective lens, it will form a hazy spot in the image plane, instead of an innitesimal point, due to diffraction
stimulated emission depletion microscopy (STED) has wide application, the intensity of depletion beam is too high that photobleaching of °uorophores and photodamage to specimens could be induced within short time.[39]
Summary
Optical microscopy is one of the key, versatile and powerful tool in scientic researches, especially in biology. Some novel far- ̄eld super-resolution °uorescence microscopies have been developed to break through the di®raction limit These microscopies include stimulated emission depletion microscopy (STED),[6] (saturated) structured illumination microscopy ((S)SIM)[7,8] based on patterned illumination, and (°uorescence) photoactivation localization microscopy ((F)PALM)[9,10] or stochastic optical reconstruction microscopy (STORM)[11] relying on single molecule localization. STED has wide application, the intensity of depletion beam is too high that photobleaching of °uorophores and photodamage to specimens could be induced within short time.[39] This property mainly limits the long-time use of STED in live samples Another commonly used far- ̄eld super-resolution microscopy is called structure illumination microscopy (SIM). Post-imaging processing may be a problem for biologists who are not familiar with the optical setup and algorithm
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