Cell Staining: Fluorescent Labelling of the Endoplasmic Reticulum

Abstract

The endoplasmic reticulum (ER) performs functions of protein and lipid synthesis and calcium regulation. Under optimal conditions, its complex structure of interconnected tubules and cisternae can be imaged by high‐resolution light microscopy. Immunofluorescence methods, which are useful in many situations, usually destroy the delicate ER structure during fixation and permeabilization.

Keywords: tubules; cisternae; dyes; applications; visualization

Figure 1.

A mouse 3T3 fibroblast cell stained with DiOC6(3). The cell was grown in culture on a glass coverslip, fixed in glutaraldehyde and stained with the dye. The staining was photographed with a conventional wide‐field fluorecence microscope using ×100 objective lens. The dye stains mitochondria and other organelles, but the most striking membranes are those of the endoplasmic reticulum (ER), which is the extensive network throughout the cell. The ER consists of interconnected tubules and cisternae (flat, planar membranes).

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References

Kline D, Mehlmann L, Fox C and Terasaki M (1999) The cortical endoplasmic reticulum (ER) of the mouse egg: localization of ER clusters in relation to the generation of repetitive calcium waves. Developmental Biology 215: 431–442.

Knebel W, Quader H and Schnepf E (1990) Mobile and immobile endoplasmic reticulum in onion bulb epidermis cells: short‐ and long‐term observations with a confocal laser scanning microscope. European Journal of Cell Biology 52: 328–340.

Koning AJ, Lum PY, Williams JM and Wright R (1993) DiOC6 staining reveals organelle structure and dynamics in living yeast cells. Cell Motility and the Cytoskeleton 25: 111–128.

Terasaki M (1998) Labeling of endoplasmic reticulum with DiOC6(3). In: Celis J (ed.) Cell Biology: A Laboratory Handbook, 2nd edn, vol. 2, pp. 501–506. Orlando, FL: Academic Press.

Terasaki M (2000) Dynamics of the ER and Golgi apparatus during early sea urchin development. Molecular Biology of the Cell 11: 897–914.

Terasaki M and Jaffe LA (1991) Organization of the sea urchin egg endoplasmic reticulum and its reorganization at fertilization. Journal of Cell Biology 114: 929–940.

Terasaki M, Song J, Wong JR, Weiss MJ and Chen LB (1984) Localization of endoplasmic reticulum in living and glutaraldehyde fixed cells with fluorescent dyes. Cell 38: 101–108.

Terasaki M, Slater NT, Fein A, Schmidek A and Reese TS (1994) A continuous cellular network of endoplasmic reticulum in cerebellar Purkinje neurons. Proceedings of the National Academy of Sciences of the USA 91: 7510–7514.

Terasaki M, Chen LB and Fujiwara K (1986) Microtubules and the endoplasmic reticulum are highly interdependent structures. Journal of Cell Biology 103: 1557–1568.

Terasaki M, Jaffe LA, Hunnicutt GR and Hammer JA III (1996) Structural change of the endoplasmic reticulum during fertilization: evidence for loss of membrane continuity using the green fluorescent protein. Developmental Biology 179: 320–328.

Waterman‐Storer CM and Salmon ED (1998) Endoplasmic reticulum membrane tubules are distributed by microtubules in living cells using three distinct mechanisms. Current Biology 8: 798–806.

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How to Cite close
Terasaki, Mark(Apr 2001) Cell Staining: Fluorescent Labelling of the Endoplasmic Reticulum. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002631]