Video-enhanced light microscopy and its applications in cell biology

1988 ◽  
Vol 89 (2) ◽  
pp. 129-150
Author(s):  
D.M. Shotton
Keyword(s):  
Image Processing ◽  
Light Microscopy ◽  
Light Microscope ◽  
Cell Biology ◽  
Living Cells ◽  
Cellular Structures ◽  
Low Contrast ◽  
Microscope Images ◽  
Microscopic Techniques

The combination of novel optical microscopic techniques with advanced video and digital image-processing technology now permits dramatic improvements in the quality of light-microscope images. Such video-enhanced light microscopy has lead to a renaissance in the applications of the light microscope for the study of living cells in two important areas: the intensification of faint fluorescence images, permitting observation of fluorescently labelled cells under conditions of very low illuminating intensity; and the enhancement of extremely low contrast images generated by minute cellular structures, so that these may be clearly seen and their normal intracellular movements recorded. Application of both these aspects of video-enhanced light microscopy have recently led to major discoveries concerning the functioning of the living cell. In this review I discuss the equipment, procedures and image-processing principles employed in these applications, and describe and illustrate some of the spectacular results that have recently been obtained.

scholarly journals A tribute to Shinya Inoue and innovation in light microscopy

2004 ◽  
Vol 165 (1) ◽  
pp. 21-26 ◽  
Author(s):  
Karen R. Dell ◽  
Ronald D. Vale
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Light Microscope ◽  
Light And Electron Microscopy ◽  
Single Molecules ◽  
Living Cells ◽  
Dynamic Processes ◽  
New Techniques

The 2003 International Prize for Biology was awarded to Shinya Inoue for his pioneering work in visualizing dynamic processes within living cells using the light microscope. He and his scientific descendants are now pushing light microscopy even further by developing new techniques such as imaging single molecules, visualizing processes in living animals, and correlating results from light and electron microscopy.

scholarly journals Review: Biofunctionalized Quantum Dots in Biology and Medicine

2009 ◽  
Vol 2009 ◽  
pp. 1-17 ◽  
Author(s):  
Sonal Mazumder ◽  
Rajib Dey ◽  
M. K. Mitra ◽  
S. Mukherjee ◽  
G. C. Das
Keyword(s):  
Quantum Dots ◽  
Optical Properties ◽  
Contrast Agents ◽  
Cell Biology ◽  
Living Cells ◽  
Fluorescence Probes ◽  
Cellular Structures ◽  
Biologically Relevant ◽  
Comprehensive Compilation

Quantum dot (QD) nanocrystals which have important optical properties, in particular, the wavelength of their fluorescence, depend strongly on their size. Colloidal QDs once dispersed in a solvent are quite interesting fluorescence probes for all types of labelling studies because of their reduced tendency to photo bleach. In this review, we will give an overview on how QDs have been used so far in cell biology. In particular, we will discuss the biologically relevant properties of QDs and focus on four topics: labeling of cellular structures and receptors with QDs, incorporation of QDs by living cells, tracking the path and the fate of individual cells using QD labels, and QDs as contrast agents. QDs are seen to be much better in terms of efficacy over radioisotopes in tracing medicine in vivo. They are rapidly being applied to existing and emerging technologies but here this review deals with a comprehensive compilation of the biological relevance of quantum dots. It covers important information from 1999 till 2008 with few from 1982 to 1997.

Image Enhancement and Restoration Methods for Underwater Images

2018 ◽  
pp. 2211-2232
Author(s):  
C. J. Prabhakar ◽  
P. U. Praveen Kumar
Keyword(s):  
Image Processing ◽  
Computer Vision ◽  
Image Enhancement ◽  
Underwater Imaging ◽  
Low Contrast ◽  
Underwater Environment ◽  
Challenging Tasks ◽  
Restoration Techniques ◽  
Restoration Algorithms

In this chapter, the authors provide an overview of state-of-the-art image enhancement and restoration techniques for underwater images. Underwater imaging is one of the challenging tasks in the field of image processing and computer vision. Usually, underwater images suffer from non-uniform lighting, low contrast, diminished color, and blurring due to attenuation and scattering of light in the underwater environment. It is necessary to preprocess these images before applying computer vision techniques. Over the last few decades, many researchers have developed various image enhancement and restoration algorithms for enhancing the quality of images captured in underwater environments. The authors introduce a brief survey on image enhancement and restoration algorithms for underwater images. At the end of the chapter, we present an overview of our approach, which is well accepted by the image processing community to enhance the quality of underwater images. Our technique consists of filtering techniques such as homomorphic filtering, wavelet-based image denoising, bilateral filtering, and contrast equalization, which are applied sequentially. The proposed method increases better image visualization of objects which are captured in underwater environment compared to other existing methods.

Image Enhancement and Restoration Methods for Underwater Images

2014 ◽  
pp. 93-110
Author(s):  
C. J. Prabhakar ◽  
P. U. Praveen Kumar
Keyword(s):  
Image Processing ◽  
Computer Vision ◽  
Image Enhancement ◽  
Underwater Imaging ◽  
Low Contrast ◽  
Underwater Environment ◽  
Challenging Tasks ◽  
Restoration Techniques ◽  
Restoration Algorithms

In this chapter, the authors provide an overview of state-of-the-art image enhancement and restoration techniques for underwater images. Underwater imaging is one of the challenging tasks in the field of image processing and computer vision. Usually, underwater images suffer from non-uniform lighting, low contrast, diminished color, and blurring due to attenuation and scattering of light in the underwater environment. It is necessary to preprocess these images before applying computer vision techniques. Over the last few decades, many researchers have developed various image enhancement and restoration algorithms for enhancing the quality of images captured in underwater environments. The authors introduce a brief survey on image enhancement and restoration algorithms for underwater images. At the end of the chapter, we present an overview of our approach, which is well accepted by the image processing community to enhance the quality of underwater images. Our technique consists of filtering techniques such as homomorphic filtering, wavelet-based image denoising, bilateral filtering, and contrast equalization, which are applied sequentially. The proposed method increases better image visualization of objects which are captured in underwater environment compared to other existing methods.

scholarly journals A quick guide to light microscopy in cell biology

2016 ◽  
Vol 27 (2) ◽  
pp. 219-222 ◽  
Author(s):  
Kurt Thorn
Keyword(s):  
Light Microscopy ◽  
Fluorescent Probes ◽  
Cell Biology ◽  
Living Cells ◽  
Diverse Range

Light microscopy is a key tool in modern cell biology. Light microscopy has several features that make it ideally suited for imaging biology in living cells: the resolution is well-matched to the sizes of subcellular structures, a diverse range of available fluorescent probes makes it possible to mark proteins, organelles, and other structures for imaging, and the relatively nonperturbing nature of light means that living cells can be imaged for long periods of time to follow their dynamics. Here I provide a brief introduction to using light microscopy in cell biology, with particular emphasis on factors to be considered when starting microscopy experiments.

At the cutting edge: applications and perspectives of laser nanosurgery in cell biology

2012 ◽  
Vol 393 (4) ◽  
pp. 235-248 ◽  
Author(s):  
Paolo Ronchi ◽  
Stefan Terjung ◽  
Rainer Pepperkok
Keyword(s):  
Drosophila Melanogaster ◽  
Danio Rerio ◽  
Cell Biology ◽  
Living Cells ◽  
Model Systems ◽  
Cellular Structures ◽  
Intracellular Space ◽  
Whole Cells ◽  
Potential Applications ◽  
Number Of Publications

Abstract Laser-mediated nanosurgery has become popular in the last decade because of the previously unexplored possibility of ablating biological material inside living cells with sub-micrometer precision. A number of publications have shown the potential applications of this technique, ranging from the dissection of sub-cellular structures to surgical ablations of whole cells or tissues in model systems such as Drosophila melanogaster or Danio rerio. In parallel, the recent development of micropatterning techniques has given cell biologists the possibility to shape cells and reproducibly organize the intracellular space. The integration of these two techniques has only recently started yet their combination has proven to be very interesting. The aim of this review is to present recent applications of laser nanosurgery in cell biology and to discuss the possible developments of this approach, particularly in combination with micropattern-mediated endomembrane organization.

scholarly journals Advantages of Simultaneous Imaging Using an Atomic Force Microscope Integrated with an Inverted Light Microscope

2011 ◽  
Vol 19 (6) ◽  
pp. 22-29
Author(s):  
W. Travis Johnson
Keyword(s):  
Atomic Force Microscopy ◽  
Light Microscopy ◽  
Scanning Probe Microscopy ◽  
Light Microscope ◽  
Cell Biology ◽  
Numerical Aperture ◽  
Scanning Probe ◽  
Force Microscopy ◽  
Simultaneous Imaging ◽  
Atomic Force

Atomic Force Microscopy (AFM) permits measurements on biological samples below the limits of light microscopy resolution under physiological environments and other controlled conditions. Consequently, AFM has become an increasingly valuable technique in cell biology. One of the most exciting advances in AFM instrumentation has been its integration with the light microscope. This permits investigators to take advantage of the power and utility of light microscopy and scanning probe microscopy simultaneously. In combining a light microscope with an AFM, scanner components must be specifically designed so that they do not adversely impact the light microscope's optical imaging capabilities. For example, an AFM-ILM (inverted light microscope) hybrid system should be fully compatible with the highest quality, off-the-shelf 0.50–0.55 NA numerical aperture (NA) OEM objectives and condensers.

scholarly journals Video Enhancement Utilizing Old and Low Contrast

2019 ◽  
Vol 54 (6) ◽  
Author(s):  
Ameen A. Noor ◽  
Ziad M. Abood ◽  
Ali Shakir Mahmood
Keyword(s):  
Image Processing ◽  
Video Clip ◽  
Disease Diagnosis ◽  
Improve Quality ◽  
Video Enhancement ◽  
Still Images ◽  
Low Contrast ◽  
Long Time ◽  
Image Density

It has been relied upon and is still found in the fields of scientific research, especially astronomy, medicine (for accurate disease diagnosis), biology, archeology, and industry on video and still images. The low accuracy and quality of some videos are often due to a poor lens type or angle, which may be due to a lack of photographic experience, or because of the older sections, which can be affected by the coolness of the surrounding perimeter. This research was completed using simple methods of processing based on using a program to convert video to individual images, then a number of image processing operations to improve quality, and finally re-assemble the images to the video more accurately than the original and in our own way. The proposed process consists of several steps: cutting the video clip into a set of images, performing various operations, such as using the contrast filter first, discovering the edges, smoothing the image, and improving image density prior to assembly. We finally assemble the images back into clips. This has been the process we used on many of the films affected by noise, or damaged for a long time, and has proven our ability to improve the quality of the video.

Selective Staining of Acid Mucopolysaccharides By Ruthenium Red

1968 ◽  
Vol 26 ◽  
pp. 38-39
Author(s):  
J. H. Luft
Keyword(s):  
Electron Microscope ◽  
Light Microscopy ◽  
Light Microscope ◽  
Osmium Tetroxide ◽  
Single Cells ◽  
Ruthenium Red ◽  
Collagen Fibers ◽  
Selective Staining ◽  
Pectic Substances ◽  
Solid Tissues

Ruthenium red is one of the few completely inorganic dyes used to stain tissues for light microscopy. This novelty is enhanced by ignorance regarding its staining mechanism. However, its continued usefulness in botany for demonstrating pectic substances attests to selectivity of some sort. Whether understood or not, histochemists continue to be grateful for small favors.Ruthenium red can also be used with the electron microscope. If single cells are exposed to ruthenium red solution, sufficient mass can be bound to produce observable density in the electron microscope. Generally, this effect is not useful with solid tissues because the contrast is wasted on the damaged cells at the block surface, with little dye diffusing more than 25-50 μ into the interior. Although these traces of ruthenium red which penetrate between and around cells are visible in the light microscope, they produce negligible contrast in the electron microscope. However, its presence can be amplified by a reaction with osmium tetroxide, probably catalytically, to be easily visible by EM. Now the density is clearly seen to be extracellular and closely associated with collagen fibers (Fig. 1).

Application of FRAP and digitized fluorescence microscopy to problems in cell membrane dynamics

1988 ◽  
Vol 46 ◽  
pp. 118-119
Author(s):  
K. Jacobson ◽  
A. Ishihara ◽  
B. Holifield ◽  
F. Zhang
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Extended Period ◽  
Dynamic Structure ◽  
Living Cells ◽  
Membrane Dynamics ◽  
Molecular Cell Biology ◽  
Image Averaging ◽  
Single Living Cells ◽  
Single Living

Our laboratory is concerned with understanding the dynamic structure of the plasma membrane with particular reference to the movement of membrane constituents during cell locomotion. In addition to the standard tools of molecular cell biology, we employ both fluorescence recovery after photo- bleaching (FRAP) and digitized fluorescence microscopy (DFM) to investigate individual cells. FRAP allows the measurement of translational mobility of membrane and cytoplasmic molecules in small regions of single, living cells. DFM is really a new form of light microscopy in that the distribution of individual classes of ions, molecules, and macromolecules can be followed in single, living cells. By employing fluorescent antibodies to defined antigens or fluorescent analogs of cellular constituents as well as ultrasensitive, electronic image detectors and video image averaging to improve signal to noise, fluorescent images of living cells can be acquired over an extended period without significant fading and loss of cell viability.

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