Cytocentrifugation is a common technique for the capture of cells on microscopic slides. It usually requires a special cytocentrifuge or cytorotor and cassettes. In the study presented here, we tested the new concept of cytocentrifugation based on the threaded connection of the lid and the sample holder to ensure an adjustable flow of solutions through the filters and the collection of the filtered solutions in the reservoir during centrifugation. To test this concept, we developed a device for the preparation of cell samples on circular coverslips. The device was tested for the capture and sample processing of both eukaryotic and prokaryotic cells, cell nuclei, and mitochondria for microscopy analysis including image cytometry. Moreover, an efficient procedure was developed for capturing formaldehyde-fixed cells on non-treated coverslips without cell drying. The results showed that the tested arrangement enables the effective capture and processing of all of the tested samples and the developed device represents an inexpensive alternative to common cytocentrifuges, as only the paper filter is consumed during sample processing, and no special centrifuge, cytorotor, or cassette is necessary. As no additional system of solution removal is required during sample staining, the tested concept also facilitates the eventual automation of the staining procedure.
Biocompatibility testing of new materials is often performed in vitro by measuring the growth rate of mammalian cancer cells in time-lapse images acquired by phase contrast microscopes. The growth rate is measured by tracking cell coverage, which requires an accurate automatic segmentation method. However, cancer cells have irregular shapes that change over time, the mottled background pattern is partially visible through the cells and the images contain artifacts such as halos. We developed a novel algorithm for cell segmentation that copes with the mentioned challenges. It is based on temporal differences of consecutive images and a combination of thresholding, blurring, and morphological operations. We tested the algorithm on images of four cell types acquired by two different microscopes, evaluated the precision of segmentation against manual segmentation performed by a human operator, and finally provided comparison with other freely available methods. We propose a new, fully automated method for measuring the cell growth rate based on fitting a coverage curve with the Verhulst population model. The algorithm is fast and shows accuracy comparable with manual segmentation. Most notably it can correctly separate live from dead cells.
- MeSH
- biologické jevy MeSH
- cytologické techniky metody přístrojové vybavení MeSH
- fluorescenční mikroskopie * MeSH
- fluorescenční protilátková technika MeSH
- fluorescenční spektrometrie metody přístrojové vybavení MeSH
- mikrobiologické techniky * metody přístrojové vybavení MeSH
- mikroskopie skenující sondou * metody přístrojové vybavení MeSH
- nanotechnologie MeSH
- MeSH
- cytologické techniky metody přístrojové vybavení MeSH
- fixace tkání metody trendy MeSH
- imunohistochemie metody trendy MeSH
- kryoprezervace ekonomika trendy MeSH
- lidé MeSH
- nádory mozku genetika patologie MeSH
- nukleové kyseliny analýza MeSH
- zalévání tkání do parafínu metody trendy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- souhrny MeSH
- MeSH
- cytologické techniky metody přístrojové vybavení využití MeSH
- hematologické testy metody přístrojové vybavení využití MeSH
- krevní nemoci diagnóza krev MeSH
- krevní obraz metody přístrojové vybavení využití MeSH
- lidé MeSH
- nemoci kostní dřeně diagnóza MeSH
- počet leukocytů metody přístrojové vybavení využití MeSH
- počítače trendy využití MeSH
- tekutiny a sekrety tělesné MeSH
- vyšetřování kostní dřeně metody přístrojové vybavení využití MeSH
- Check Tag
- lidé MeSH
Electromagnetic fields generated by living cells have been experimentally investigated in the past 3 decades; however, the results are often inconsistent. In this paper we discuss some technical aspects of such challenging experiments, a brief review of which is also included. Special attention is paid to the sensor with respect to the power available from a cell and the power needed to excite the macroscopic measurement devices. We drew the conclusion that the nanoelectronic approach should be used.
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- MeSH
- cytodiagnostika metody trendy využití MeSH
- cytologické techniky metody přístrojové vybavení trendy využití MeSH
- průtoková cytometrie metody přístrojové vybavení trendy využití MeSH
- Publikační typ
- abstrakt z konference MeSH
- sborníky MeSH
- Konspekt
- Buněčná biologie. Cytologie
- NLK Obory
- cytologie, klinická cytologie
