Two Pathways in Pollen Protoplast Culture: Cell Divisions and Tube Growth

Exposure of cells to low sublethal but mitosis-arresting doses of vinblastine sulfate (Velban) results in the initial arrest of cells in mitosis followed by their subsequent return to an “interphase“-like stage. A large number of these cells reform their nuclear membranes and form large multimicronucleated cells, some containing as many as 25 or more micronuclei (1). Formation of large multinucleate cells is also caused by cytochalasin, by causing the fusion of daughter cells at the end of an otherwise .normal cell division (2). By the repetition of this process through subsequent cell divisions, large cells with 6 or more nuclei are formed.

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A Method for Electron Microscopic Study of Tissue Culture Cell Monolayers Grown in Tubes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060672 ◽

1967 ◽

Vol 25 ◽

pp. 132-133

Author(s):

R. Stephens ◽

G. Schidlovsky ◽

S. Kuzmic ◽

P. Gaudreau

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Light Microscopy ◽

Cell Sheet ◽

Culture Cell ◽

Tissue Culture Cell ◽

Electron Microscopic ◽

Cell Sheets ◽

Morphological Alterations ◽

Culture Cells

The usual method of scraping or trypsinization to detach tissue culture cell sheets from their glass substrate for further pelletization and processing for electron microscopy introduces objectionable morphological alterations. It is also impossible under these conditions to study a particular area or individual cell which have been preselected by light microscopy in the living state.Several schemes which obviate centrifugation and allow the embedding of nondetached tissue culture cells have been proposed. However, they all preserve only a small part of the cell sheet and make use of inverted gelatin capsules which are in this case difficult to handle.We have evolved and used over a period of several years a technique which allows the embedding of a complete cell sheet growing at the inner surface of a tissue culture roller tube. Observation of the same cell by light microscopy in the living and embedded states followed by electron microscopy is performed conveniently.

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Application of stereological analysis of cell volume to isolated myocytes in culture with and without adrenergic innervation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010015962x ◽

1990 ◽

Vol 48 (3) ◽

pp. 416-417

Author(s):

Jane K. Rosenthal ◽

Dianne L. Atkins ◽

William J. Marvin ◽

Penny A. Krumm

Keyword(s):

Cardiac Myocytes ◽

Structural Changes ◽

Three Dimensional ◽

Adrenergic Innervation ◽

Culture Cell ◽

Serial Sections ◽

Newborn Rats ◽

Primary Culture Cell ◽

Biological Studies ◽

Cell Volumes

To comprehend structural changes in cardiac myocytes accompanying adrenergic innervation, it is essential that a three dimensional analysis be performed. To date, biological studies which utilize stereological methods have been limited to cells in tissue and in organs. Our laboratory has utilized current stereological techniques for measuring absolute volumes of individual myocytes in primary culture. Cell volumes are calculated for two distinct groups of cells at 96 hours in culture: isolated myocytes and myocytes innervated with adrenergic neurons (Figure 1).Cardiac myocytes are cultured from the ventricular apices of newborn rats. Cells are plated directly onto tissue culture dishes with or without preplated explants from the paravertebral thoracolumbar sympathetic chain. On day four cultures are photographed and marked for one-to-one cell location. Following conventional fixation and embeddment in eponate-12, the cells are relocated and mounted for microtomy. The cells are completely sectioned at 120nm in their parallel orientation to the surface of the dish (Figure 2). Serial sections are collected on formvar coated slotted grids and are recorded in sequence.

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