scholarly journals Intermittent swimming in live sea urchin sperm.

1980 ◽  
Vol 84 (1) ◽  
pp. 1-12 ◽  
Author(s):  
B H Gibbons
Keyword(s):  
Sea Urchin ◽  
Divalent Cations ◽  
Dark Field ◽  
Proximal Region ◽  
Ionophore A23187 ◽  
Quiescent State ◽  
Sodium Barbital ◽  
Dark Field Microscopy ◽  
Short Period ◽  
Live Sperm

Sperm of the sea urchin Tripneustes gratilla repeatedly start and stop swimming when suspended in seawater and observed by dark-field microscopy. While in the quiescent state, which usually lasts about a second, the sperm assume s shape resembling a cane, with a sharp bend of approximately 3.4 rad in the proximal region of the flagellum and very little curvature in the rest of the flagellum except for a slight curve near the tip. The occurrence of quiescence requires the presence of at least 2 mM Ca2+ in the seawater, and the percentage of sperm quiescent at any one time increases substantially when the sperm are illuminated with blue light. With intense illumination, close to 100% of the sperm become quiescent, and this percentage decreases gradually to approximately 0.3% over a 10(4)-fold decrease in light intensity. An increased concentration of K+ in the seawater also increases the percentage of quiescence, with a majority of the sperm being quiescent in seawater containing 80 mM KCl. The induction of quiescence by light or by increased KCl is completely inhibited by 10 micrometers chlorpromazine, and approximately 90% inhibited by 1 mM procaine or sodium barbital. Sperm treated with the divalent-cation ionophore A23187 swim quite normally, although for a relatively short period, in artificial seawater lacking divalent cations, but are abruptly arrested upon addition of 0.04--0.2 mM free Ca2%. The flagellar waveform of these arrested sperm is almost identical to that of light-induced quiescence in the live sperm. The results support the hypothesis that quiescence is induced by a rise in intracellular Ca2%, perhaps as a consequence of a membrane depolarization, and that it is similar to the arrest response in cilia.

scholarly journals Conformational change in the outer doublet microtubules from sea urchin sperm flagella.

1979 ◽  
Vol 81 (2) ◽  
pp. 355-360 ◽  
Author(s):  
T Miki-Noumura ◽  
R Kamiya
Keyword(s):  
Sea Urchin ◽  
Conformational Changes ◽  
Helical Structure ◽  
Dark Field ◽  
Ion Concentration ◽  
Wave Form ◽  
Buffer System ◽  
Left Handed ◽  
Dark Field Microscopy ◽  
Hemicentrotus Pulcherrimus

Dark-field microscopy with a high-powered light source revealed that the outer doublet microtubules (DMTs) from sea urchin (Pseudocentrotus depressus and Hemicentrotus pulcherrimus) sperm flagella assume helically coiled configurations (Miki-Noumura, T., and R. Kamiya. 1976. Exp. Cell Res. 97: 451.). We report here that the DMTs change shape when the pH or Ca-ion concentration is changed. The DMTs assumed a left-handed helical shape with a diameter of 3.7 +/- 0.5 micron and a pitch of 2.8 +/- 0.7 micron at pH 7.4 in the presence of 0.1 mM CaCl2, 1 mM MgSO4, and 10 mM Tris-HCl. When the pH was raised to 8.3, the helical diameter and pitch decreased to 2.1 +/- 0.1 micron and 1.3 +/- 0.3 micron, respectively. This transformation was a rapid and reversible process and was completed within 1 min. Between pH 7.2 and 8.3, the DMTs assumed intermediate shapes. When the Ca-ion concentration was depleted with EGTA, the helical structure became significantly larger in both pitch and diameter. For instance, the diameter was 3.8 +/- 0.4 micron at pH 8.3 in the presence of 1 mM EGTA and 2 mM MgSO4. Using a Ca-buffer system, we obtained results which suggested that this Ca-induced transformation took place at a Ca concentration of approximately 10(-7) M. These results were highly reproducible. The conformational changes in the DMT may play some role in the bending wave form of flagellar movement.

scholarly journals Calcium-induced quiescence in reactivated sea urchin sperm.

1980 ◽  
Vol 84 (1) ◽  
pp. 13-27 ◽  
Author(s):  
B H Gibbons ◽  
I R Gibbons
Keyword(s):  
Sea Urchin ◽  
Proximal Region ◽  
Bend Angle ◽  
Elevated Concentration ◽  
Bending Waves ◽  
Close Relationship ◽  
Gentle Agitation ◽  
Live Sperm ◽  
Triton X ◽  
Asymmetrical Bending

Sperm flagella of the sea urchin Tripneustes gratilla beat with asymmetrical bending waves after demembranation with Triton X-100 in the presence of EGTA and reactivation at pH 8.1 with 1 mM ATP in the presence of 2 mM MgSO4. Addition of 0.1--0.2 mM free Ca2+ to these reactivated sperm induces 70--95% of them to become quiescent. This quiescence can be reversed by reduction of the free Ca2% concentration with EGTA, or by dilution to reduce the MgATP2- concentration below 0.3 mM. The quiescent waveform is characterized by a sharp principal bend of approximately 5.6 rad in the proximal region of the flagellum, a slight reverse bend in the midregion that averages approximately 0.3 rad, and a principal bend of approximately 1.1 rad in the tip. The quiescent sperm are highly fragile mechanically, and disruption, including microtubule sliding, occurs spontaneously at a slow rate upon standing or immediately upon gentle agitation. Mild digestion by trypsin causes a gradual appearance of normal, symmetrical flagellar beating. Addition of increasing concentrations of vanadate to quiescent sperm causes a graded decrease in the proximal bend angle, with 50 micrometers vanadate reducing it to approximately 2.6 rad. In the presence of 0.1 mM free Ca2% and 10 micrometers vanadate, a characteristic, crescented stationary bend is induced in the demembranated sperm, without intermediate oscillatory beating, by the addition of either 0.1 or 1 mM ATP. In the absence of vanadate, these two concentrations of ATP produce asymmetric beating and quiescence, respectively. The results support the hypothesis that quiescence in live sperm is induced by an elevated concentration of intracellular Ca2%. In addition, they demonstrate that bending can occur in flagella in which oscillatory beating is inhibited and emphasize the close relationship between asymmetric beating and quiescence.

Elastic Scattering in Electron Microscopy

1973 ◽  
Vol 31 ◽  
pp. 252-253
Author(s):  
J. Langmore ◽  
M. Isaacson ◽  
J. Wall ◽  
A. V. Crewe
Keyword(s):  
Electron Microscopy ◽  
Elastic Scattering ◽  
Cross Section ◽  
Quantum Noise ◽  
Dark Field ◽  
Elastic Cross Section ◽  
Biological Molecules ◽  
Dark Field Microscopy ◽  
Field Systems ◽  
Effects Of Radiation

High resolution dark field microscopy is becoming an important tool for the investigation of unstained and specifically stained biological molecules. Of primary consideration to the microscopist is the interpretation of image Intensities and the effects of radiation damage to the specimen. Ignoring inelastic scattering, the image intensity is directly related to the collected elastic scattering cross section, σɳ, which is the product of the total elastic cross section, σ and the eficiency of the microscope system at imaging these electrons, η. The number of potentially bond damaging events resulting from the beam exposure required to reduce the effect of quantum noise in the image to a given level is proportional to 1/η. We wish to compare η in three dark field systems.

Simulated Dark-Field Electron Micrographs of Point Defects and Organometallics

1975 ◽  
Vol 33 ◽  
pp. 200-201
Author(s):  
William Krakow
Keyword(s):  
Electron Micrographs ◽  
Point Defects ◽  
Structure Determination ◽  
Atomic Structure ◽  
Image Interpretation ◽  
Dark Field ◽  
Field Electron ◽  
Dark Field Microscopy ◽  
Dark Field Electron

Tilted beam dark-field microscopy has been applied to atomic structure determination in perfect crystals, several synthesized molecules with heavy atcm markers and in the study of displaced atoms in crystals. Interpretation of this information in terms of atom positions and atom correlations is not straightforward. Therefore, calculated dark-field images can be an invaluable aid in image interpretation.

scholarly journals Multiplexed Plasmonic Nano-Labeling for Bioimaging of Cytological Stained Samples

Cancers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 3509
Author(s):  
Paule Marcoux-Valiquette ◽  
Cécile Darviot ◽  
Lu Wang ◽  
Andrée-Anne Grosset ◽  
Morteza Hasanzadeh Kafshgari ◽  
...  
Keyword(s):  
Accurate Determination ◽  
Cell Types ◽  
Fine Needle Biopsy ◽  
Dark Field ◽  
Imaging Method ◽  
New Methods ◽  
Formalin Fixed Paraffin ◽  
Dark Field Microscopy ◽  
Formalin Fixed Paraffin Embedded

Reliable cytopathological diagnosis requires new methods and approaches for the rapid and accurate determination of all cell types. This is especially important when the number of cells is limited, such as in the cytological samples of fine-needle biopsy. Immunoplasmonic-multiplexed- labeling may be one of the emerging solutions to such problems. However, to be accepted and used by the practicing pathologists, new methods must be compatible and complementary with existing cytopathology approaches where counterstaining is central to the correct interpretation of immunolabeling. In addition, the optical detection and imaging setup for immunoplasmonic-multiplexed-labeling must be implemented on the same cytopathological microscope, not interfere with standard H&E imaging, and operate as a second easy-to-use imaging method. In this article, we present multiplex imaging of four types of nanoplasmonic markers on two types of H&E-stained cytological specimens (formalin-fixed paraffin embedded and non-embedded adherent cancer cells) using a specially designed adapter for SI dark-field microscopy. The obtained results confirm the effectiveness of the proposed optical method for quantitative and multiplex identification of various plasmonic NPs, and the possibility of using immunoplasmonic-multiplexed-labeling for cytopathological diagnostics.

scholarly journals Hyperspectral-enhanced dark field analysis of individual and collective photo-responsive gold–copper sulfide nanoparticles

Nanoscale ◽  
2021 ◽  
Author(s):  
Paula Zamora-Perez ◽  
Beatriz Pelaz ◽  
Dionysia Tsoutsi ◽  
Mahmoud G. Soliman ◽  
Wolfgang J. Parak ◽  
...  
Keyword(s):  
Physicochemical Properties ◽  
Real Time ◽  
Copper Sulfide ◽  
Dark Field ◽  
Field Analysis ◽  
Dark Field Microscopy

Hyperspectral-enhanced dark field microscopy to correlate Au/CuS NPs’ changes in their physicochemical properties induced by cellular environments with their functionality as photothermal probes by tracking their scattering profile evolution in real time.

scholarly journals Dark-field microscopy enhance visibility of CD31 endothelial staining

2020 ◽  
Vol 64 (3) ◽  
Author(s):  
Eva Jennische ◽  
Stefan Lange ◽  
Ragnar Hultborn
Keyword(s):  
Phase Contrast ◽  
Scattered Light ◽  
Dark Field ◽  
Light Scatter ◽  
Endothelial Lining ◽  
Microscopy Technique ◽  
Dark Field Microscopy ◽  
Dark Field Illumination ◽  
Normal Human ◽  
Intense Light

A simple dark field microscopy technique was used for visualization of blood vessels in normal human renal tissues and carcinoma. Phase contrast condenser ring apt for high power objectives was combined with a 10x objective in order to create a dark field illumination of the specimens examined. The endothelial lining of the vessels had been stained by using CD31 monoclonal antibodies combined with conventional peroxidase immunohistochemistry. The final DAB addition used for this technique induced an intense light scatter in the dark field microscope. This scattered light originating from the endothelial lining made the walls of the bright vessels easily detectable from the dark background.

Effect of imposed head vibration on the stability and waveform of flagellar beating in sea urchin spermatozoa

1991 ◽  
Vol 156 (1) ◽  
pp. 63-80 ◽  
Author(s):  
C. Shingyoji ◽  
I. R. Gibbons ◽  
A. Murakami ◽  
K. Takahashi
Keyword(s):  
Sea Urchin ◽  
Beat Frequency ◽  
Distal Region ◽  
Proximal Region ◽  
Bend Angle ◽  
Vibration Frequencies ◽  
Sliding Velocity ◽  
Additional Energy ◽  
The Stability ◽  
Cyclic Changes

The heads of live spermatozoa of the sea urchin Hemicentrotus pulcherrimus were held by suction in the tip of a micropipette mounted on a piezoelectric device and vibrated either laterally or axially with respect to the head axis. Within certain ranges of frequency and amplitude, lateral vibration of the pipette brought about a stable rhythmic beating of the flagella in the plane of vibration, with the beat frequency synchronized to the frequency of vibration [Gibbons et al. (1987), Nature 325, 351–352]. The sperm flagella, with an average natural beat frequency of 48 Hz, showed stable beating synchronized to the pipette vibration over a range of 35–90 Hz when the amplitude of vibration was about 20 microns or greater. Vibration frequencies below this range caused instability of the beat plane, often associated with irregularities in beat frequency. Frequencies above about 90 Hz caused irregular asymmetrical flagellar beating with a marked decrease in amplitude of the propagated bends and a skewing of the flagellar axis towards one side; the flagella often stopped in a cane shape. In flagella that were beating stably under imposed vibration, the wavelength was reduced at higher frequencies and increased at lower frequencies. When the beat frequency was equal to or lower than the natural beat frequency, the apparent time-averaged sliding velocity of axonemal microtubules, obtained as twice the product of frequency and bend angle, decreased with beat frequency in both the proximal and distal regions of the flagella. However, at vibration frequencies above the natural beat frequency, the sliding velocity increased with frequency only in the proximal region of the flagellum and remained essentially unchanged in more distal regions. This apparent limit to the velocity of sliding in the distal region may represent an inherent limit in the intrinsic velocity of active sliding, while the faster sliding observed in the proximal region may be a result of passive sliding or elastic distortion of the microtubules induced by the additional energy supplied by the vibrating pipette. Axial vibration with frequencies either close to or twice the natural beat frequency induced cyclic changes in the waveform, compressing and expanding the bends in the proximal region, but did not affect bends in the distal region or alter the beat frequency.

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