A simple re-embedding method for the preparation of testicular cell cultures for light and electron microscopy

3D cell cultures, in particular organoids, recapitulate tissue architectures and are emerging model systems to investigate healthy or diseased tissues and organs. Understanding the complex cellular sociology of organoids requires the integration of diverse imaging modalities covering different spatial and temporal resolutions. Here, we present an integrated multi-scale 3D imaging approach that traverses from millimeter-scale live-cell light microscopy to nanoscale volume electron microscopy. Performing the 3D cell culture in a single carrier amenable to all imaging steps provides a continuum resolution view of the same organoid. Our approach allows us to follow organoid growth, probe their morphology with fluorescent markers, identify cells of interest and analyze their ultrastructure. We demonstrate this workflow on mouse and human derived 3D cell cultures. The continuum resolution imaging pipeline is thus suited to foster both basic and translational organoid research by simultaneously exploiting the advantages of light and electron microscopy.

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Immune Electron Microscopy (IEM) of a Canine Adeno-Associated Virus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051293 ◽

1974 ◽

Vol 32 ◽

pp. 256-257

Author(s):

Gunter F. Thomas ◽

M. David Hoggan

Keyword(s):

Electron Microscopy ◽

Cell Cultures ◽

Complement Fixation ◽

Hepatitis Virus ◽

Wide Distribution ◽

Immune Electron Microscopy ◽

Adeno Associated Virus ◽

Virus Like Particle ◽

Dog Kidney ◽

Green Monkeys

In 1968, Sugimura and Yanagawa described a small 25 nm virus like particle in association with the Matsuda strain of infectious canine hepatitis virus (ICHV). Domoto and Yanagawa showed that this particle was dependent on ICHV for its replication in primary dog kidney cell cultures (PDK) and was resistant to heating at 70°C for 10 min, and concluded that it was a canine adeno-associated virus (CAAV). Later studies by Onuma and Yanagawa compared CAAV with the known human serotypes (AAV 1, 2, 3) and AAV-4, known to be associated with African Green Monkeys. Using the complement fixation (CF) test, they found that CAAV was serologically related to AAV-3 and had wide distribution in the dog population of Japan.

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The Morphology of Vacuolated Cells in the Liver Following Administration of Vitamin A.

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072484 ◽

1973 ◽

Vol 31 ◽

pp. 408-409

Author(s):

Odell T. Minick ◽

Hidejiro Yokoo ◽

Fawzia Batti

Keyword(s):

Electron Microscopy ◽

Body Weight ◽

Vitamin A ◽

Light And Electron Microscopy ◽

Liver Cells ◽

Prominent Feature ◽

Vascular Space ◽

Vacuolated Cell ◽

Vacuolated Cells ◽

Vacuolated Cytoplasm

Vacuolated cells in the liver of young rats were studied by light and electron microscopy following the administration of vitamin A (200 units per gram of body weight). Their characteristics were compared with similar cells found in untreated animals.In rats given vitamin A, cells with vacuolated cytoplasm were a prominent feature. These cells were found mostly in a perisinusoidal location, although some appeared to be in between liver cells (Fig. 1). Electron microscopy confirmed their location in Disse's space adjacent to the sinusoid and in recesses between liver cells. Some appeared to be bordering the lumen of the sinusoid, but careful observation usually revealed a tenuous endothelial process separating the vacuolated cell from the vascular space. In appropriate sections, fenestrations in the thin endothelial processes were noted (Fig. 2, arrow).

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Morphological Examination of a Herpes-type Virus Indigenous for Tree Shrews

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100067790 ◽

1970 ◽

Vol 28 ◽

pp. 160-161

Author(s):

J. P. Brunschwig ◽

R. M. McCombs ◽

R. Mirkovic ◽

M. Benyesh-Melnick

Keyword(s):

Electron Microscopy ◽

Soft Tissue ◽

Chick Embryo ◽

Soft Tissue Sarcoma ◽

Cell Cultures ◽

Tree Shrew ◽

Type Virus ◽

Morphological Examination ◽

Tree Shrews ◽

Embryo Cells

A new virus, established as a member of the herpesvirus group by electron microscopy, was isolated from spontaneously degenerating cell cultures derived from the kidneys and lungs of two normal tree shrews. The virus was found to replicate best in cells derived from the homologous species. The cells used were a tree shrew cell line, T-23, which was derived from a spontaneous soft tissue sarcoma. The virus did not multiply or did so poorly for a limited number of passages in human, monkey, rodent, rabbit or chick embryo cells. In the T-23 cells, the virus behaved as members of the subgroup B of herpesvirus, in that the virus remained primarily cell associated.

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Scanning Electron Microscopy and Electron Microprobe Analysis of Malignant Cell Cultures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100561 ◽

1968 ◽

Vol 26 ◽

pp. 164-165

Author(s):

R. I. Johnsson-Hegyeli ◽

A. F. Hegyeli ◽

D. K. Landstrom ◽

W. C. Lane

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Cell Cultures ◽

Electron Microprobe ◽

Microprobe Analysis ◽

Electron Microprobe Analysis ◽

Three Dimensional ◽

Malignant Cell ◽

Interference Microscopy ◽

Scanning Electron

Last year we reported on the use of reflected light interference microscopy (RLIM) for the direct color photography of the surfaces of living normal and malignant cell cultures without the use of replicas, fixatives, or stains. The surface topography of living cells was found to follow underlying cellular structures such as nuceloli, nuclear membranes, and cytoplasmic organelles, making possible the study of their three-dimensional relationships in time. The technique makes possible the direct examination of cells grown on opaque as well as transparent surfaces. The successful in situ electron microprobe analysis of the elemental composition and distribution within single tissue culture cells was also reported.This paper deals with the parallel and combined use of scanning electron microscopy (SEM) and the two previous techniques in a study of living and fixed cancer cells. All three studies can be carried out consecutively on the same experimental specimens without disturbing the cells or their structural relationships to each other and the surface on which they are grown. KB carcinoma cells were grown on glass coverslips in closed Leighto tubes as previously described. The cultures were photographed alive by means of RLIM, then fixed with a fixative modified from Sabatini, et al (1963).

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Control of Autogenous Vein Grafts Prior to Reimplantation, By Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010009378x ◽

1972 ◽

Vol 30 ◽

pp. 106-107

Author(s):

John H. L. Watson ◽

John L. Swedo ◽

M. Vrandecic

Keyword(s):

Electron Microscopy ◽

Light And Electron Microscopy ◽

Physiological Saline ◽

Experimental Conditions ◽

Vein Grafts ◽

Arterial Blood ◽

Saphenous Veins ◽

Autogenous Vein ◽

Control Of Parameters ◽

Post Implantation

The ambient temperature and the nature of the storage fluids may well have significant effects upon the post-implantation behavior of venus autografts. A first step in the investigation of such effects is reported here. Experimental conditions have been set which approximate actual operating room procedures. Saphenous veins from dogs have been used as models in the experiments. After removal from the dogs the veins were kept for two hours under four different experimental conditions, viz at either 4°C or 23°C in either physiological saline or whole canine arterial blood. At the end of the two hours they were prepared for light and electron microscopy. Since no obvious changes or damage could be seen in the veins by light microscopy, even with the advantage of tissue specific stains, it was essential that the control of parameters for successful grafts be set by electron microscopy.

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Ultrastructure of two neoplasms in culture compared with original biopsies

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100110623 ◽

1984 ◽

Vol 42 ◽

pp. 50-51

Author(s):

Joseph M. Harb ◽

James T. Casper ◽

Vlcki Piaskowski

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Biopsy Specimen ◽

Cultured Cells ◽

Light And Electron Microscopy ◽

Human Tumor ◽

Soft Agar ◽

Human Tumor Cells ◽

Morphological Distinction ◽

Tumor Types

The application of tissue culture and the newer methodologies of direct cloning and colony formation of human tumor cells in soft agar hold promise as valuable modalities for a variety of diagnostic studies, which include morphological distinction between tumor types by electron microscopy (EM). We present here two cases in which cells in culture expressed distinct morphological features not apparent in the original biopsy specimen. Evaluation of the original biopsies by light and electron microscopy indicated both neoplasms to be undifferentiated sarcomas. Colonies of cells propagated in soft agar displayed features of rhabdomyoblasts in one case, and cultured cells of the second biopsy expressed features of Ewing's sarcoma.

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