Procedure for in situ Embedding of Cell Cultures on Plastic Petri Dish

1971 ◽  
Keyword(s):  
Cell Cultures ◽  
Petri Dish ◽  
Plastic Petri Dish ◽  
In Situ Embedding

Rotary Beveling for In Situ Thin-Section Microscopy of Cell Cultures

1981 ◽  
Vol 39 ◽  
pp. 550-551
Author(s):  
Dean A. Handley ◽  
Jack T. Alexander ◽  
Shu Chien
Keyword(s):  
Cell Growth ◽  
Cell Cultures ◽  
Molecular Interactions ◽  
Convenient Method ◽  
Petri Dish ◽  
Simple Method ◽  
Thin Section Microscopy ◽  
Thin Sectioning ◽  
Organic Fluids

In situ preparation of cell cultures for ultrastructural investigations is a convenient method by which fixation, dehydration and embedment are carried out in the culture petri dish. The in situ method offers the advantage of preserving the native orientation of cell-cell interactions, junctional regions and overlapping configurations. In order to section after embedment, the petri dish is usually separated from the polymerized resin by either differential cryo-contraction or solvation in organic fluids. The remaining resin block must be re-embedded before sectioning. Although removal of the petri dish may not disrupt the native cellular geometry, it does sacrifice what is now recognized as an important characteristic of cell growth: cell-substratum molecular interactions. To preserve the topographic cell-substratum relationship, we developed a simple method of tapered rotary beveling to reduce the petri dish thickness to a dimension suitable for direct thin sectioning.

scholarly journals Lab-Tek Chamber Slide for TEM Prep: A Simple, Rapid, and Reliable Protocol for In Situ Embedding Monolayer Cell Cultures in Epoxy and LR White Resin

2008 ◽  
Vol 16 (1) ◽  
pp. 38-41
Author(s):  
Gang Ning
Keyword(s):  
Cell Cultures ◽  
Cellular Interaction ◽  
Mechanical Trauma ◽  
Special Importance ◽  
Monolayer Cell ◽  
Differentiated Cells ◽  
Microcentrifuge Tube ◽  
Transmission Electron ◽  
In Situ Embedding

Preparation for TEM of resin-embedded monolayer cell cultures usually requires scraping cells from the culture substrate before or after primary fixation, which disturbs the monolayer and often results in mechanical trauma to the cells. After harvest, the cells are centrifuged and processed as a pellet in a microcentrifuge tube through a prolonged procedure of post-fixation, dehydration, infiltration, and finally embedding to ensure a “well done” sample block for subsequent sectioning, staining, and observing under a transmission electron microscope. Other disadvantages to this method include the loss of cellular orientation and information on cellular interaction, as well as difficulty in collecting a large quantity of cells to form a sizable pellet for processing, which is of special importance when samples are differentiated cells and neurons cultured in low density. In an alternative method, cells can be seeded on either glass or plastic cover slips or Petri dishes, and then processed and embedded directly as a monolayer.

The Fine Structure of Rat Granulosa Cell Cultures Correlated with Progestin Secretion

1973 ◽  
Vol 31 ◽  
pp. 552-553
Author(s):  
T. M. Crisp ◽  
F.R. Denys
Keyword(s):  
Fine Structure ◽  
Granulosa Cell ◽  
Cell Cultures ◽  
Single Injection ◽  
Surrounding Medium ◽  
Petri Dish ◽  
Female Rats ◽  
Vaginal Smear ◽  
Immature Female ◽  
Serum Gonadotropin

The purpose of this paper is to present observations on the fine structure of rat granulosa cell cultures grown in the presence of an adenohypophyseal explant and to correlate the morphology of these cells with progestin secretion. Twenty-six day old immature female rats were given a single injection of 5 IU pregnant mares serum gonadotropin (PMS) in order to obtain ovaries with large vesicular follicles. At 66 hrs. post-PMS administration (estrus indicated by vaginal smear cytology), the ovaries were removed and placed in a petri dish containing medium 199 and 100 U penicillin/streptomycin (P/S)/ml. Under a 20X magnification dissecting microscope, some 5-8 vesicular follicles/ovary were punctured and the granulosa cells were expressed into the surrounding medium. The cells were transferred to centrifuge tubes and spun down at 1000 rpm for 5 mins.

scholarly journals Electrical impedance measurement system to assess in-situ experiments on epithelia under ELF magnetic fields exposure

2021 ◽  
Vol 19 (3) ◽  
pp. 217-226
Author(s):  
G. Domínguez ◽  
E. Cardiel ◽  
J.L Reyes ◽  
E. Sánchez ◽  
P.R. Hernández
Keyword(s):  
Impedance Spectroscopy ◽  
Magnetic Fields ◽  
Cell Cultures ◽  
Electrical Impedance ◽  
Controlled Environment ◽  
Spectroscopy Technique ◽  
In Situ Experiments ◽  
Impedance Spectroscopy Technique

Purpose: The development of an electric impedance meter based on the impedance spectroscopy technique, for in vitro and in situ experimentation, with cellular epithelia submitted to extremely low frequency magnetic fields in a controlled environment. Unlike other reported systems, a strength of the one presented here is that it avoids the influence of external factors on the experiment. Materials and methods: The designed system employs the electrical impedance values obtained by the impedance spectroscopy technique to determine the parameters of the simple equivalent electrical model of a cellular monolayer. The Madin-Darby Canine Kidney (MDCK) cell cultures were used as subjects of study in the experimental protocol. Results: The validation was carried out by comparing the transepithelial electrical impedance data of the cell cultures obtained with the developed system and those of the Cellzscope® commercial system used as the standard. Non-significant differences were obtained. Conclusion: It was confirmed that the developed system provides reliable values of transepithelial electrical impedance to experiment with cell cultures and take advantage of the controlled environment to reduce the effects of experimental management.

scholarly journals A Novel Flat-embedding Method to Prepare Ultrathin Cryosections from Cultured Cells in Their In Situ Orientation

2002 ◽  
Vol 50 (8) ◽  
pp. 1067-1080 ◽  
Author(s):  
Viola Oorschot ◽  
Heidi de Wit ◽  
Wim G. Annaert ◽  
Judith Klumperman
Keyword(s):  
Electron Microscopy ◽  
Quantitative Method ◽  
Cultured Cells ◽  
Petri Dish ◽  
Ultrastructural Level ◽  
Immunogold Labeling ◽  
Polarized Cells ◽  
Ultrathin Cryosections ◽  
Flat Embedding

Immunogold labeling of ultrathin cryosections provides a sensitive and quantitative method to localize proteins at the ultrastructural level. An obligatory step in the routine preparation of cryosections from cultured cells is the detachment of cells from their substrate and subsequent pelleting. This procedure precludes visualization of cells in their in situ orientation and hampers the study of polarized cells. Here we describe a method to sample cultured cells from a petri dish or coverslip by embedding them in a 12% gelatin slab. Subsequently, sections can be prepared in parallel or perpendicular to the plane of growth. Our method extends the cryosectioning technique to applications in studying polarized cells and correlative light–electron microscopy.

The viability of animal cell cultures in bioreactors: Can it be estimated online by using in situ microscopy?

2010 ◽  
Vol 45 (2) ◽  
pp. 288-291 ◽  
Author(s):  
J.S. Guez ◽  
J.Ph. Cassar ◽  
F. Wartelle ◽  
P. Dhulster ◽  
H. Suhr
Keyword(s):  
Cell Cultures ◽  
Animal Cell ◽  
Animal Cell Cultures

scholarly journals Laboratory Evaluation of a Neem Product Against DBM Larvae, 1995

1997 ◽  
Vol 22 (1) ◽  
pp. 409-409
Author(s):  
R.F.L. Mau ◽  
L. R. Gusukuma-Minuto
Keyword(s):  
Petri Dish ◽  
Seedling Stage ◽  
Laboratory Evaluation ◽  
Complete Coverage ◽  
Plastic Petri Dish ◽  
True Leaf ◽  
Laboratory Colony ◽  
Treated Plant ◽  
Head Cabbage

Abstract Treatments were evaluated using the leaf dip method. Head cabbage was seeded in community pots. Each pot containing approximately 10 cabbage plants in the 5 true leaf seedling stage was inverted and dipped in a test insecticide mix for about 30 sees for complete coverage. The dipped plants were allowed to air dry. For each dip, 1 liter insecticide mix was prepared based on field rate concentrations of 100 gal/acre. Leaves from treated plants were detached and placed in a ventilated plastic petri dish. DBM larvae from a laboratory colony that originated from individuals collected from a cabbage field at Kula, Hawaii and Kamuela, Hawaii were used. Ten late 2nd instars were placed on each leaf. Fresh leaves from the original treated plant were added every two days. The number of dead larvae was counted at 24-hour intervals. Larvae were recorded as dead when there was no movement when probed.

In situ quantification of microcarrier animal cell cultures using near-infrared spectroscopy

2010 ◽  
Vol 45 (8) ◽  
pp. 1427-1431 ◽  
Author(s):  
Emma Petiot ◽  
Patrick Bernard-Moulin ◽  
Thierry Magadoux ◽  
Cécile Gény ◽  
Hervé Pinton ◽  
...  
Keyword(s):  
Infrared Spectroscopy ◽  
Cell Cultures ◽  
Near Infrared Spectroscopy ◽  
Near Infrared ◽  
Animal Cell ◽  
Animal Cell Cultures
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