Skip to main content
Log in

Interaction of molecular probes with living cells and tissues. Part 2

A structure-activity analysis of mitochondrial staining by cationic probes, and a discussion of the synergistic nature of image-based and biochemical approaches

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

Cultured rat fibroblasts were exposed to 41 cationic fluorescent probes of very varied hydrophilicity/lipophilicity. Outcome of probe-cell interaction fell into one of the following categories: probe failed to enter the cells; probe accumulated on cell surfaces; probe accumulated in mitochondria, and/or in other intracellular regions. The observations were analysed using a Simplistic Chinese Box (SCB) approach, and the following conclusions were reached. It was the hydrophilic probes which failed to enter cells, whilst extremely lipophilic probes were retained on the cell surfaces. Only the slightly lipophilic cationic probes were permeant, and accumulated in mitochondria. Using the probes log P values to model hydrophilicity/lipophilicity, effective cationic mitochondrial stains can be specified numerically so: 0<log P probe<+5. This SCB model was used to rationalise a variety of earlier observations on the action of mitochondrial probes. The applicability of the SCB approach to integrate image-based and biochemical investigations was demonstrated by using the action of chlorpromazine on mitochondrial action as a case example.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alberts B, Bray D, Lewis J, Raff M, Roberts K, Watson JD (1983) Molecular biology of the cell. Garland Publishing, New York London, p 486

    Google Scholar 

  • Berger M (1957) Metabolic reactivity of brain and liver mitochondria towards chlorpromazine. J Neurochem 2:30–36

    Google Scholar 

  • Brenner S (1953) Supravital staining of mitochondria with pheno-safranin dyes. Biochim Biophys Acta 11:480–486

    Google Scholar 

  • Chen LB (1989) The fluorescent labelling of mitochondria-A review. Methods Cell Biol 29:103–123

    Google Scholar 

  • Clark G, Kasten FH (1983) History of staining, 3rd. edn. Williams and Wilkins, Baltimore

    Google Scholar 

  • Cowden RR, Curtis SK (1986) In vitro (supravital) fluorescence cytochemistry. Basic Appl Histochem 30:7–40

    Google Scholar 

  • Cowdry EV (1916) The general functional significance of mitochondria. Am J Anat 9:423–446

    Google Scholar 

  • DuBuy HG, Showacre JL (1961) Selective localization of tetracycline in mitochondria of living cells. Science 133:196–197

    Google Scholar 

  • Erbrich U, Berthold Th, Zimmerman HW (1982) Effects of acridine dyes on the ultrastructure of mitochondria in HeLa and LM cells. Histochemistry 76:211–218

    Google Scholar 

  • Hansch C, Leo AJ (1979) Substituent constants for correlation analysis in chemistry and biology. Wiley, New York, chapter 4

    Google Scholar 

  • Haugland RP (1989) Handbook of fluorescent probes and research chemicals. Molecular Probes, Eugene, p 125

    Google Scholar 

  • Hetherington DC (1936) Pinacyanol as a supra-vital mitochondrial stain for blood. Stain Technol 11:153–154

    Google Scholar 

  • Horobin RW, Rashid F (1990) Interaction of molecular probes with living cells and tissues. 1. Some general mechanistic proposals, making use of the simplistic chinese-box model. Histochemistry 94:205–209

    Google Scholar 

  • Johnson LV, Walsh ML, Chen LB (1980) Localisation of mitochondria in living cells with Rhodamine 123. Proc Natl Acad Sci USA 77:990–994

    Google Scholar 

  • Johnson LV, Walsh ML, Bockus BJ, Chen LB (1981) Monitoring of relative mitochondrial membrane potential in living cells by fluorescent microscopy. J Cell Biol 88:526–535

    Google Scholar 

  • Lewis MR, Lewis WH (1919) Mitochondria (and other cytoplasmic organelles) in tissue cultures. Am J Anat 17(3):339–401

    Google Scholar 

  • Payne JN, Cooper JD, MacKeown ST, Horobin RW (1987) A temperature controlled chamber to allow observation and measurement of uptake of fluorochromes into live cells. J Microsc 147:329–335

    Google Scholar 

  • Proctor GB, Horobin RW (1985) A widely applicable analytical system for biological stains: reverse-phase thin layer chromatography. Stain Technol 60:1–6

    Google Scholar 

  • Röding J, Naujok A, Zimmerman HW (1986) Effects of ethidium bromide, tetramethylethidium bromide and betaine B on the ultrastructure of HeLa cell mitochondria in situ. Histochemistry 85:215–222

    Google Scholar 

  • Simpson ME (1921) Vital staining of human blood with special reference to the separation of the monocytes. Univ California Public Anat 1:1–9

    Google Scholar 

  • Strickland EH, Benson AA (1960) Neuron activation paper chromatography analysis of phosphatides in mammalian cell fractions. Arch Biochem Biophys 88:344–348

    Google Scholar 

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

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rashid, F., Horobin, R.W. Interaction of molecular probes with living cells and tissues. Part 2. Histochemistry 94, 303–308 (1990). https://doi.org/10.1007/BF00266632

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00266632

Keywords

Navigation