Fourier-filtering based size-encoded images for label-free tracking of sub-cellular organelles in neurons

2019 ◽  
Author(s):  
Mohammad Naser ◽  
Erin Kelly ◽  
Corey Ditmars ◽  
Rene S. Schloss ◽  
Bonnie N. Firestein ◽  
...  
Keyword(s):  
Label Free ◽  
Fourier Filtering ◽  
Cellular Organelles

scholarly journals Label free 3D analysis of organelles in living cells by refractive index shows pre-mitotic organelle spinning in mammalian stem cells

2018 ◽  
Author(s):  
Patrick A. Sandoz ◽  
Christopher Tremblay ◽  
Sebastien Equis ◽  
Sorin Pop ◽  
Lisa Pollaro ◽  
...  
Keyword(s):  
Refractive Index ◽  
Lipid Droplets ◽  
Living Cells ◽  
Live Cells ◽  
3D Analysis ◽  
Label Free ◽  
Long Period ◽  
Microscopy Method ◽  
First Time ◽  
Cellular Organelles

AbstractHolo-tomographic microscopy (HTM) is a label-free non-phototoxic microscopy method reporting the fine changes of a cell’s refractive indexes (RI) in 3D. By combining HTM with epifluorescence, we demonstrate that cellular organelles such as Lipid droplets and mitochondria show a specific RI signature that distinguishes them with high resolution and contrast. We further show that HTM allows to follow in unprecedented ways the dynamics of mitochondria, lipid droplets as well as that of endocytic structures in live cells over long period of time, which led us to observe to our knowledge for the first time a global organelle spinning occurring before mitosis.

A complex network of “snake-like tubules” revealed by the NADPase cytochemical reaction in the acinar cell of pancreas

1984 ◽  
Vol 42 ◽  
pp. 666-667
Author(s):  
A.R. Beaudoin ◽  
G. Grondin ◽  
A. Lord ◽  
M. Pelletier
Keyword(s):  
Acinar Cell ◽  
Lead Acetate ◽  
Ultrastructural Localization ◽  
Sodium Acetate Buffer ◽  
Zymogen Granules ◽  
Sprague Dawley ◽  
Dense Bodies ◽  
Sprague Dawley Rats ◽  
Cytochemical Reaction ◽  
Cellular Organelles

We have recently described the ultrastructural localization of NADPase activity in the exocrine pancreas of rat. The enzyme was found in the intermediate saccules of the Golgi apparatus, in dense bodies and lysosomes but was absent from zymogen granules. A very intense reaction was noticed in a peculiar structure which was termed “Snake-Like Tubule” (SLT). The purposes of the present study were firstly to delineate SLT distribution in the acinar cell and secondly to define any possible relationship or association with other cellular organelles.NADPase cytochemical reaction was performed on the pancreas of adult Sprague Dawley rats. Small lobules were excised and fixed for 50 min, at 4°C, in 2% glutaraldehyde buffered with 0.1M cacodylate at pH 7.2. Lobules were rinsed several times with the same buffer containing 570 sucrose and cut with a Mcllwayn tissue chopper. Sections were washed several times with buffer and incubated for 2 hr at 37°C in the following medium: 4mM NADPH; 40mM sodium acetate buffer, pH 5.0; 4mM lead acetate and 5% sucrose.

Application of cross correlation to HREM images of surfaces

1987 ◽  
Vol 45 ◽  
pp. 754-755
Author(s):  
D. E. Luzzi ◽  
L. D. Marks ◽  
M. I. Buckett
Keyword(s):  
Cross Correlation ◽  
A Priori ◽  
Matrix Material ◽  
Correlation Method ◽  
Accurate Knowledge ◽  
Complex Image ◽  
Fourier Filtering ◽  
The Matrix ◽  
Low Pass ◽  
Data Reduction Technique

As the HREM becomes increasingly used for the study of dynamic localized phenomena, the development of techniques to recover the desired information from a real image is important. Often, the important features are not strongly scattering in comparison to the matrix material in addition to being masked by statistical and amorphous noise. The desired information will usually involve the accurate knowledge of the position and intensity of the contrast. In order to decipher the desired information from a complex image, cross-correlation (xcf) techniques can be utilized. Unlike other image processing methods which rely on data massaging (e.g. high/low pass filtering or Fourier filtering), the cross-correlation method is a rigorous data reduction technique with no a priori assumptions.We have examined basic cross-correlation procedures using images of discrete gaussian peaks and have developed an iterative procedure to greatly enhance the capabilities of these techniques when the contrast from the peaks overlap.

Scanning Secondary Electron Microscopy of Myofibrils Using Transmission Techniques

1975 ◽  
Vol 33 ◽  
pp. 556-557
Author(s):  
M. K. Lamvik
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Carbon Films ◽  
Photographic Recording ◽  
Transmission Electron ◽  
Thin Carbon ◽  
The Common ◽  
Scanning Electron ◽  
Better Than ◽  
Cellular Organelles

When observing small objects such as cellular organelles by scanning electron microscopy, it is often valuable to use the techniques of transmission electron microscopy. The common practice of mounting and coating for SEM may not always be necessary. These possibilities are illustrated using vertebrate skeletal muscle myofibrils.Micrographs for this study were made using a Hitachi HFS-2 scanning electron microscope, with photographic recording usually done at 60 seconds per frame. The instrument was operated at 25 kV, with a specimen chamber vacuum usually better than 10-7 torr. Myofibrils were obtained from rabbit back muscle using the method of Zak et al. To show the component filaments of this contractile organelle, the myofibrils were partially disrupted by agitation in a relaxing medium. A brief centrifugation was done to clear the solution of most of the undisrupted myofibrils before a drop was placed on the grid. Standard 3 mm transmission electron microscope grids covered with thin carbon films were used in this study.

Two structural states of the Z-band in cardiac and skeletal muscle

1989 ◽  
Vol 47 ◽  
pp. 1022-1023
Author(s):  
J.P. Schroeter ◽  
M.A. Goldstein ◽  
J.P. Bretaudiere ◽  
L.H. Michael ◽  
R.L. Sass
Keyword(s):  
Skeletal Muscle ◽  
Cross Section ◽  
Real Space ◽  
Contractile State ◽  
Contour Maps ◽  
False Color ◽  
Grey Scale ◽  
Fourier Filtering ◽  
Cardiac Muscles ◽  
Enhancement Algorithm

We have recently established the existence of two structural states of the Z band lattice in cross section in cardiac as well as in skeletal muscle. The two structural states are related to the contractile state of the muscle. In skeletal muscle at rest, the Z band is in the small square (ss) lattice form, but tetanized muscle exhibits the basket weave (bw) form. In contrast, unstimu- lated cardiac muscle exhibits the bw form, but cardiac muscles exposed to EGTA show the ss form.We have used two-dimensional computer enhancement techniques on digitized electron micrographs to compare each lattice form as it appears in both cardiac and skeletal muscle. Both real space averaging and fourier filtering methods were used. Enhanced images were displayed as grey-scale projections, as contour maps, and in false color.There is only a slight difference between the lattices produced by the two different enhancement techniques. Thus the information presented in these images is not likely to be an artifact of the enhancement algorithm.

Label-free, mass-sensitive single-molecule imaging using interferometric scattering microscopy

2020 ◽  
Author(s):  
Nikolas Hundt
Keyword(s):  
Single Molecule ◽  
Cellular Systems ◽  
Single Molecule Imaging ◽  
Label Free ◽  
Measurement Sensitivity ◽  
Mass Distributions ◽  
Quantitative Measurements ◽  
Contrast Mechanism ◽  
Cellular Components ◽  
Scattering Signal

Abstract Single-molecule imaging has mostly been restricted to the use of fluorescence labelling as a contrast mechanism due to its superior ability to visualise molecules of interest on top of an overwhelming background of other molecules. Recently, interferometric scattering (iSCAT) microscopy has demonstrated the detection and imaging of single biomolecules based on light scattering without the need for fluorescent labels. Significant improvements in measurement sensitivity combined with a dependence of scattering signal on object size have led to the development of mass photometry, a technique that measures the mass of individual molecules and thereby determines mass distributions of biomolecule samples in solution. The experimental simplicity of mass photometry makes it a powerful tool to analyse biomolecular equilibria quantitatively with low sample consumption within minutes. When used for label-free imaging of reconstituted or cellular systems, the strict size-dependence of the iSCAT signal enables quantitative measurements of processes at size scales reaching from single-molecule observations during complex assembly up to mesoscopic dynamics of cellular components and extracellular protrusions. In this review, I would like to introduce the principles of this emerging imaging technology and discuss examples that show how mass-sensitive iSCAT can be used as a strong complement to other routine techniques in biochemistry.

A microfluidic device with integrated impedance detection for λ-DNA

2003 ◽  
Vol 773 ◽  
Author(s):  
Myung-Il Park ◽  
Jonging Hong ◽  
Dae Sung Yoon ◽  
Chong-Ook Park ◽  
Geunbae Im
Keyword(s):  
Microfluidic Device ◽  
Electrochemical Impedance ◽  
Direct Detection ◽  
Full Potential ◽  
Label Free ◽  
Buffer Solution ◽  
Detection Systems ◽  
Significant Difference ◽  
Detection Of Biomolecules ◽  
Impedance Magnitude

AbstractThe large optical detection systems that are typically utilized at present may not be able to reach their full potential as portable analysis tools. Accurate, early, and fast diagnosis for many diseases requires the direct detection of biomolecules such as DNA, proteins, and cells. In this research, a glass microchip with integrated microelectrodes has been fabricated, and the performance of electrochemical impedance detection was investigated for the biomolecules. We have used label-free λ-DNA as a sample biomolecule. By changing the distance between microelectrodes, the significant difference between DW and the TE buffer solution is obtained from the impedance-frequency measurements. In addition, the comparison for the impedance magnitude of DW, the TE buffer, and λ-DNA at the same distance was analyzed.

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