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Imaging Biological Samples with Optical Microscopy01:18

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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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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Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...
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Total Internal Reflection Fluorescence Microscopy01:05

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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.
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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...
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Related Experiment Video

Updated: May 4, 2026

Biomolecular Imaging of Cellular Uptake of Nanoparticles using Multimodal Nonlinear Optical Microscopy
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Multimodal Nonlinear Optical Microscopy.

Shuhua Yue1, Mikhail N Slipchenko1, Ji-Xin Cheng1

  • 1Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907.

Laser & Photonics Reviews
|December 20, 2013
PubMed
Summary
This summary is machine-generated.

Multimodal nonlinear optical (NLO) imaging combines techniques like CARS microscopy to study complex biological tissues. This approach enhances understanding of diseases and enables development of advanced NLO imaging modalities.

Keywords:
CARSFWMNLO microscopySHGSRSTPEFbiomedicalconfocal Ramanmultimodalnanomaterialphotothermaltransient absorption

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Area of Science:

  • Biomedical Imaging
  • Microscopy
  • Optical Physics

Background:

  • Multimodal nonlinear optical (NLO) imaging leverages the specificity of different NLO modalities for comprehensive studies of biological tissues.
  • Combining techniques like multiphoton fluorescence, second harmonic generation, and coherent anti-Stokes Raman scattering (CARS) addresses diverse biological questions.

Purpose of the Study:

  • To review approaches for creating multimodal NLO imaging.
  • To discuss the development of advanced NLO modalities on a CARS microscope platform.
  • To highlight applications in biological and biomedical research.

Main Methods:

  • Integration of multiphoton fluorescence, second harmonic generation, and CARS.
  • Utilizing CARS microscopy as a platform for novel NLO modalities (e.g., electronic-resonance-enhanced four-wave mixing, stimulated Raman scattering, pump-probe microscopy).

Main Results:

  • Successful application of multimodal NLO imaging to lipid metabolism, cancer, cardiovascular disease, and skin biology.
  • Demonstration of CARS microscope's versatility in developing advanced NLO techniques.
  • Broad applicability of these imaging methods across various research areas.

Conclusions:

  • Multimodal NLO imaging offers powerful capabilities for investigating complex biological systems.
  • The CARS microscope serves as a valuable platform for advancing NLO imaging technologies.
  • These advanced imaging techniques are crucial for progress in biological and biomedical research.