In situ prolyl oligopeptidase activity assay in neural cell cultures

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.

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Author Correction: An in situ activity assay for lysyl oxidases

Communications Biology ◽

10.1038/s42003-021-02655-4 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Huilei Wang ◽

Alan Poe ◽

Lydia Pak ◽

Kavitha Nandakumar ◽

Sandeep Jandu ◽

...

Keyword(s):

Activity Assay ◽

Lysyl Oxidases

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

Journal of Applied Research and Technology ◽

10.22201/icat.24486736e.2021.19.3.1693 ◽

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.

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