Fluorescence cell imaging and manipulation using conventional halogen lamp microscopy.

PloS One
Kazuo YamagataTeruhiko Wakayama

Abstract

Technologies for vitally labeling cells with fluorescent dyes have advanced remarkably. However, to excite fluorescent dyes currently requires powerful illumination, which can cause phototoxic damage to the cells and increases the cost of microscopy. We have developed a filter system to excite fluorescent dyes using a conventional transmission microscope equipped with a halogen lamp. This method allows us to observe previously invisible cell organelles, such as the metaphase spindle of oocytes, without causing phototoxicity. Cells remain healthy even after intensive manipulation under fluorescence observation, such as during bovine, porcine and mouse somatic cell cloning using nuclear transfer. This method does not require expensive epifluorescence equipment and so could help to reduce the science gap between developed and developing countries.

References

Jan 1, 1972·Journal of Clinical Pathology·G V Heimer, C E Taylor
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Apr 1, 1995·Biology of Reproduction·Y Kimura, R Yanagimachi
Sep 5, 2003·Biology of Reproduction·Mito Kanatsu-ShinoharaTakashi Shinohara
Oct 17, 2007·The Journal of Reproduction and Development·Satoshi Kishigami, Teruhiko Wakayama
Mar 24, 2009·The Journal of Reproduction and Development·Kazuo YamagataTeruhiko Wakayama
Dec 10, 2009·The Journal of Cell Biology·Yoko Hayashi-TakanakaHiroshi Kimura
Mar 6, 2010·The Journal of Reproduction and Development·Nguyen Van ThuanTeruhiko Wakayama

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Citations

Nov 14, 2013·Cellular Reprogramming·Domenico IusoPasqualino Loi
Sep 4, 2015·Journal of Biosciences·Guido A Drexler, Miguel J Ruiz-Gomez

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Methods Mentioned

BETA
fluorescence microscopy
fluorescence-activated cell sorting
genetic modification
conventional
imaging technique
Cesarean section

Software Mentioned

Olympus

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