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Related Concept Videos

Two-Dimensional Microscopy in Microbiology01:29

Two-Dimensional Microscopy in Microbiology

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...
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...
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.
Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth
07:28

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth

Published on: November 24, 2017

Live cell fluorescence microscopy techniques.

Shawn A Galdeen1, Alison J North

  • 1Bio-Imaging Resource Center, The Rockefeller University, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 13, 2011
PubMed
Summary
This summary is machine-generated.

Researchers explore fluorescent tags for in vivo protein tracking, detailing critical live-cell imaging conditions. Maintaining physiological norms and minimizing phototoxicity are key for accurate cell function studies.

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Area of Science:

  • Cell Biology
  • Microscopy
  • Biotechnology

Background:

  • Fluorescent tags enable in vivo protein tracking, generating novel insights into cellular functions.
  • Advancements in microscopy methods are crucial for leveraging fluorescent tag technology.
  • Live-cell imaging requires strict control over environmental and optical parameters.

Purpose of the Study:

  • To provide an overview of essential considerations for in vivo protein tracking using fluorescent tags.
  • To highlight the importance of maintaining physiological conditions during live-cell imaging.
  • To discuss microscope design principles that optimize fluorescent tag methodologies.

Main Methods:

  • Review of current fluorescent tagging techniques for protein localization.
  • Analysis of environmental control systems for live-cell microscopy (temperature, CO2, media).
  • Evaluation of strategies to mitigate phototoxicity and photobleaching in fluorescence microscopy.

Main Results:

  • Identification of critical parameters for successful in vivo protein tracking.
  • Emphasis on the need for integrated approaches combining sample preparation and microscopy.
  • Discussion of trade-offs between signal detection and potential cellular damage.

Conclusions:

  • Optimizing live-cell imaging conditions is paramount for reliable in vivo protein studies.
  • Microscope design and experimental protocols must address phototoxicity and physiological stability.
  • This overview serves as a guide for researchers utilizing fluorescent tags in cell biology.