scholarly journals THE ASSOCIATION OF A CLASS OF SALTATORY MOVEMENTS WITH MICROTUBULES IN CULTURED CELLS

1970 ◽  
Vol 45 (2) ◽  
pp. 334-354 ◽  
Author(s):  
Jerome J. Freed ◽  
Marcia M. Lebowitz
Keyword(s):  
Electron Microscopy ◽  
Acid Phosphatase ◽  
Phase Contrast ◽  
Cultured Cells ◽  
Time Lapse ◽  
Carbon Particles ◽  
Time Lapse Microscopy ◽  
Use Of Time ◽  
Cytoplasmic Microtubules ◽  
Histochemical Examination

Particulate structures in the cytoplasm of HeLa and other cultured cells in interphase undergo rapid individual linear displacements (long saltatory movements, LSM). By the use of time-lapse microscopy to locate saltating particles prior to fixation and histochemical examination of the cells, structures of several kinds have been shown to move in this manner. Elements that show LSM include lysosomes, pinosomes, ingested carbon particles, lipoidal granules, and unidentified particles that appear as bright objects in positive phase contrast. The pattern of movement of the particles suggests the presence of linear guiding elements radially disposed from the cytocenter (centriole region). The participation of microtubules in these movements is inferred from the observation that LSM cease after treatment with drugs which depolymerize microtubules, i.e., colchicine, Vinblastine, and podophyllin. The directions of the microtubules in the cytoplasm of HeLa cells found by electron microscopy are consistent with the aster-like configuration predicted from study of LSM. Further support for this arrangement of cytoplasmic microtubules is provided by light microscope observations of colchicine-sensitive radial arrays of acid phosphatase granules in the cytoplasm of some cell lines.

Phase-contrast and electron-microscopic observations on a membranous complex in primary spermatocytes of the mouse

1975 ◽  
Vol 18 (1) ◽  
pp. 1-17
Author(s):  
A. Pleshkewych ◽  
L. Levine
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Phase Contrast ◽  
Cultured Cells ◽  
Light And Electron Microscopy ◽  
Cytoplasmic Inclusion ◽  
Electron Microscopic ◽  
Secondary Spermatocyte ◽  
Living Mouse ◽  
Circular Contour

A prominent cytoplasmic inclusion present in living mouse primary spermatocytes has been observed by both light and electron microscopy. It began to form at prometaphase and continued to increase in thickness and length as the cells developed. By metaphase it was a distinct sausage-shaped boundary that enclosed a portion of the cytoplasm between the spindle and the cell membrane. At the end of metaphase, the inclusion reached its maximum length. At telophase, it was divided between the daughter secondaries. The inclusion persisted as a circular contour in the interphase secondary spermatocyte. Electron microscopy of the same cultured cells that were previously observed with light microscopy revealed that the inclusion was a distinctive formation of membranes. It consisted of agranular cisternae and vesicles, and was therefore a membranous complex. Many of the smaller vesicles in the membranous complex resembled those found in the spindle. The cisternae in the membranous complex were identical to the cisternal endoplasmic reticulum of interphase primary spermatocytes. Nevertheless, the organization of vesicles and cisternae into the membranous complex was unique for the primaries in division stages, since such an organization was not present in their interphase stages.

scholarly journals Use of time-lapse microscopy to visualize rapid movement of the replication origin region of the chromosome during the cell cycle in Bacillus subtilis

1998 ◽  
Vol 28 (5) ◽  
pp. 883-892 ◽  
Author(s):  
Chris D. Webb ◽  
Peter L. Graumann ◽  
Jason A. Kahana ◽  
Aurelio A. Teleman ◽  
Pamela A. Silver ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Bacillus Subtilis ◽  
Replication Origin ◽  
Time Lapse ◽  
Time Lapse Microscopy ◽  
Rapid Movement ◽  
Use Of Time ◽  
Origin Region

scholarly journals THE CIRCUMFUSION SYSTEM FOR MULTIPURPOSE CULTURE CHAMBERS

1968 ◽  
Vol 39 (2) ◽  
pp. 430-450 ◽  
Author(s):  
George G. Rose ◽  
M. Kumegawa ◽  
M. Cattoni
Keyword(s):  
Phase Contrast ◽  
Cultured Cells ◽  
Morphological Characteristics ◽  
Time Lapse ◽  
Newborn Mouse ◽  
Contrast Microscopy ◽  
Free Environment ◽  
Parenchymal Cells

The circumfusion system is a complex in vitro pumping unit incorporating 12 multipurpose culture chambers through which a serum-supplemented fluid nutrient is recirculated at a rate of 4.5 ml/min per chamber. This system was used to study the differentiative responses of fetal and newborn mouse liver explants placed in the serum-free environment formed between the sheets of unperforated cellophane and cover glasses of the chambers. Hepatocytes (parenchymal cells) were discernible in 3–5 days. They retained many of their features of differentiation in the circumfusion system for more than 120 days of cultivation. The living morphological characteristics of the hepatocytes were studied by phase-contrast microscopy (direct viewing and time-lapse cinemicrography) and by special cytochemical staining. Electron micrographs were made of both fresh liver specimens and the cultured cells. Comparisons of the cultured parenchymal cells with their in vivo progenitors showed a remarkable preservation of their differentiated state.

scholarly journals Time-Lapse Microscopy of Streptomyces coelicolor Growth and Sporulation

2008 ◽  
Vol 74 (21) ◽  
pp. 6774-6781 ◽  
Author(s):  
Vinod Jyothikumar ◽  
Emma J. Tilley ◽  
Rashmi Wali ◽  
Paul R. Herron
Keyword(s):  
Protein Trafficking ◽  
Growth And Development ◽  
Fluorescent Protein ◽  
Imaging System ◽  
Streptomyces Coelicolor ◽  
Time Lapse ◽  
Time Lapse Microscopy ◽  
Use Of Time ◽  
Aerial Hyphae ◽  
Green Fluorescent

ABSTRACT Bacteria from the genus Streptomyces are among the most complex of all prokaryotes; not only do they grow as a complex mycelium, they also differentiate to form aerial hyphae before developing further to form spore chains. This developmental heterogeneity of streptomycete microcolonies makes studying the dynamic processes that contribute to growth and development a challenging procedure. As a result, in order to study the mechanisms that underpin streptomycete growth, we have developed a system for studying hyphal extension, protein trafficking, and sporulation by time-lapse microscopy. Through the use of time-lapse microscopy we have demonstrated that Streptomyces coelicolor germ tubes undergo a temporary arrest in their growth when in close proximity to sibling extension sites. Following germination, in this system, hyphae extended at a rate of ∼20 μm h−1, which was not significantly different from the rate at which the apical ring of the cytokinetic protein FtsZ progressed along extending hyphae through a spiraling movement. Although we were able to generate movies for streptomycete sporulation, we were unable to do so for either the erection of aerial hyphae or the early stages of sporulation. Despite this, it was possible to demonstrate an arrest of aerial hyphal development that we suggest is through the depolymerization of FtsZ-enhanced green fluorescent protein (GFP). Consequently, the imaging system reported here provides a system that allows the dynamic movement of GFP-tagged proteins involved in growth and development of S. coelicolor to be tracked and their role in cytokinesis to be characterized during the streptomycete life cycle.

Dynamic Surface Features of Cultured Cells Visualized by Combining Cinephotomicrography and SEM

1975 ◽  
Vol 33 ◽  
pp. 516-517
Author(s):  
Bruce Wetzel ◽  
Gary Jones ◽  
Edwina Westbrook ◽  
Cecil Fox ◽  
Katherine Sanford
Keyword(s):  
Cell Growth ◽  
Phase Contrast ◽  
Cultured Cells ◽  
Fetal Bovine Serum ◽  
Time Lapse ◽  
Distilled Water ◽  
Dynamic Surface ◽  
Surface Features ◽  
Fetal Bovine ◽  
And Behavior

The surface morphology of cells grown to subconfluent monolayers can be remarkably diverse and changeable. To study the dynamics of these surface features, cell growth and behavior can be observed by time-lapse cinematography without noticeably perturbing the culture,and the same cells reexamined by SEM1.Mouse fibroblasts of nonneoplastic and neoplastic lines derived from a common single cell were trypsinized, then grown in NCTC-135 with 10% filtered fetal bovine serum in a 10% CO2 atmosphere for 48 hours at 37°C on glass coverslips within sealed glass flasks. During this period a field of cells was filmed at one frame/minute with phase contrast optics (80X). Cells were fixed by the addition of an equal volume of 6% phosphate-buffered glutaraldehyde for 90 minutes at 37°C, washed with distilled water, dehydrated through ethanol to amyl acetate, dried by the CO2 critical point method, and vacuum coated with carbon and gold-palladium.

3-D analysis of cellular lipid metabolism: Reconstruction of the tubular network

1992 ◽  
Vol 50 (1) ◽  
pp. 924-925
Author(s):  
W.G. Jerome ◽  
B.F. McEwen ◽  
L.K. Minor ◽  
G.H. Rothblat ◽  
J.M. Glick
Keyword(s):  
Electron Microscopy ◽  
Smooth Muscle ◽  
Acid Phosphatase ◽  
Smooth Muscle Cells ◽  
Lipid Accumulation ◽  
Cultured Cells ◽  
Muscle Cells ◽  
Stereo Pair ◽  
Secondary Lysosomes ◽  
Relationship Of

A predominant feature of early atherosclerotic lesions is the presence of macrophages and smooth muscle cells in which large quantities of lipid have accumulated. This lipid occurs within both small, cytoplasmic, cholesterol ester droplets and large, lipid-swollen secondary lysosomes. Time resolved studies suggest lysosomal engorgement promotes the disease progression. Using cultured cells we have shown that lysosomal engorgement in smooth muscle cells is accompanied by the elaboration of an extensive network of tubules. The tubules are rich in acid hydrolases and appear to be an extension of the trans Golgi network. As part of our effort to understand the causes and significance of lysosomal lipid accumulation we have undertaken to characterize the extent and development of the tubular network. The tortuous nature of the network makes it difficult to visualize by conventional thin section electron microscopy. Thus, intermediate voltage electron microscopy of thick (1-3 um) sections in which the tubules have been contrasted using acid phosphatase cytochemistry has been essential to these studies. Cultured rabbit smooth muscle cells were incubated with sonicated lipid droplets composed of 72% cholesteryl oleate, 25% triolein, and 3% phospholipid. Prior to incubation, cells had no visible lipid stores and 92% of cells showed no acid phosphatase-containing tubules. In cells with tubules, the tubules occupied less than 0.1% of the cell volume. In contrast, after 4 h of exposure to droplets 81% of cells showed lysosomal lipid accumulation with 67% exhibiting extensive, interconnecting, acid phosphatase positive tubular networks. Although stereo-pair images provided much information about the extent of the network (figure 1), the exact relationship of the network to lipid-engorged lysosomes was somewhat ambiguous due to superposition of objects. In order to clarify the relationship of network to lysosome, a tomographic reconstruction was computed from an IVEM tilt series encompassing +/− 60° in 2° increments. Images were digitized using a DAGE model 81 video camera. Rotational alignment using fiduciary markers was accomplished using the algorithm described by Lawrence. Translational alignment and weighted back-projection reconstruction were carried out as described by McEwen. Volumes were rendered using a PIXAR computer graphics workstation. The reconstruction (figure 2) revealed that extensions of the network made contact with the lipid-swollen lysosomes suggesting that the network might transfer hydrolases directly to secondary lysosomes.

scholarly journals Neuron–Glial Interactions in the Developing Cerebellum

2012 ◽  
Vol 18 (4) ◽  
pp. 742-744 ◽  
Author(s):  
Anna Dunaevsky
Keyword(s):  
Electron Microscopy ◽  
Confocal Microscopy ◽  
Brain Slices ◽  
Time Lapse ◽  
Dynamic Interactions ◽  
Time Lapse Microscopy ◽  
Developing Cerebellum

AbstractAdvances in microscopy allow one to probe the structure of neurons and their interactions with astrocytes in brain slices and in vivo at ever increasing resolution. Moreover, the dynamic interactions between the cells can be examined in live preparation. In this paper we discuss how a variety of imaging approaches: confocal microscopy, electron microscopy, and multiphoton time-lapse microscopy are employed to probe neuron glia interactions in the developing cerebellum.

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