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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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.
Atomic Force Microscopy01:08

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...

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High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
14:09

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

Published on: November 16, 2019

Ultrastable combined atomic force and total internal reflection fluorescence microscope [corrected].

H Gumpp1, S W Stahl, M Strackharn

  • 1Chair for Applied Physics and Center for NanoScience, Ludwig-Maximilians-University Munich, Amalienstr. 54, D-80799 Munich, Germany.

The Review of Scientific Instruments
|July 2, 2009
PubMed
Summary
This summary is machine-generated.

This study presents a novel hybrid atomic force microscope (AFM) and total internal reflection fluorescence (TIRF) microscope. This improved design offers enhanced mechanical stability for advanced single-molecule investigations in life sciences.

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Last Updated: Jun 22, 2026

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
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Published on: November 16, 2019

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
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Published on: May 3, 2022

Area of Science:

  • Biophysics
  • Microscopy
  • Nanotechnology

Background:

  • Combining atomic force microscopy (AFM) with other techniques like total internal reflection fluorescence (TIRF) enhances single-molecule investigations.
  • Existing hybrid AFM-TIRF systems face limitations due to mechanical instability.

Purpose of the Study:

  • To present a novel hybrid AFM-TIRF microscope design with improved mechanical stability.
  • To overcome limitations of conventional combined microscopy techniques.

Main Methods:

  • Development of a novel hybrid AFM-TIRF microscope.
  • Detailed analysis of noise spectra.
  • Comparison of different designs and operation modes.

Main Results:

  • The novel design demonstrates superior mechanical stability compared to conventional systems.
  • Simultaneous single-molecule manipulation by AFM and TIRF imaging was achieved.
  • Thorough noise spectra analysis and design comparisons were performed.

Conclusions:

  • The developed hybrid AFM-TIRF microscope offers enhanced stability for single-molecule studies.
  • This instrument opens new possibilities for life science research using combined microscopy techniques.