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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: Jun 13, 2026

Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends
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Note: Dynamic point spread function for single and multiphoton fluorescence microscopy.

Partha Pratim Mondal1, Subhra Mandal, Alberto Diaspro

  • 1Department of Instrumentation, Indian Institute of Science, Bangalore 560012, India. partha@isu.iisc.ernet.in

The Review of Scientific Instruments
|May 6, 2010
PubMed
Summary
This summary is machine-generated.

We developed a dynamic point spread function (PSF) for fluorescence microscopy that can change shape and size. This allows for adjustable imaging depth, improving nanobioimaging capabilities.

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Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Microscopy

Background:

  • Traditional fluorescence microscopy has limitations in controlling imaging depth.
  • Achieving variable excitation depths is crucial for advanced bioimaging applications.

Purpose of the Study:

  • To propose and demonstrate a dynamic point spread function (PSF) for fluorescence microscopy.
  • To enable adjustable excitation from shallow to deep regions within a sample.

Main Methods:

  • Utilized a specially designed spatial filter.
  • Dynamically altered the parameters of the spatial filter to control the PSF.
  • Demonstrated the dynamic PSF in single and multiphoton fluorescence microscopy setups.

Main Results:

  • Successfully generated a controllable PSF that can transition between localized and elongated shapes.
  • Showcased the capability for adjustable shallow-to-depth excitation during active imaging.

Conclusions:

  • The dynamic PSF offers enhanced control over imaging depth in fluorescence microscopy.
  • Potential applications include advanced nanobioimaging and in-situ cellular studies.