Skip to main content
SpringerLink
Log in

Assessment of Cell Viability in Intact Glandular Tissue in Chironomus ramosus using Dye-exclusion and Colorimetric Assays

  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Conventionally, dye-exclusion test for determining cell viability has been restricted only for cells in suspension in tissue culture. In this paper, salivary gland of Chironomus has been proposed as a simple tissue model system where dye-exclusion test can be reliably employed for the intact gland. We have compared suitability of commonly used vital dyes and nigrosin was found suitable for the salivary gland cells. Biochemical tests using tetrazolium salts are also commonly used for determining quantitative indices of cell viability in metabolically active cells. Ours is the first attempt to extend the same technique for the whole tissue. We standardized the conditions and prepared a protocol for MTT-based colorimetric assay suitable for the salivary gland of Chironomus. A strong correlation (r2 = 0.9893) was obtained where increasing O.D. correlated linearly with the number of live glands. We concluded that nigrosin dye-exclusion and MTT metabolic inclusion assays are suitable methods for the viability test of metabolically active intact salivary gland of Chironomus which can serve as a potential model for the assessment of cytotoxicity in future.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • P. Armitage P.S. Cranston L.C.V. Pinder (Eds) (1994) Chironomidae: Biology & Ecology of Non-biting Midges Chapman & Hall London

    Google Scholar 

  • H.L. Bank (1987) ArticleTitleAssessment of islet cell viability using fluorescent dyes Diabetologia 30 812–816 Occurrence Handle10.1007/BF00275748 Occurrence Handle1:STN:280:BieC3M3lsF0%3D Occurrence Handle3322907

    Article  CAS  PubMed  Google Scholar 

  • T. Bernas J. Dubrucki (2002) ArticleTitleMitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1 and NAO mitochondrial fluorescent probes Cytometry 47 236–242 Occurrence Handle10.1002/cyto.10080 Occurrence Handle1:CAS:528:DC%2BD38XjtFSktb0%3D Occurrence Handle11933013

    Article  CAS  PubMed  Google Scholar 

  • A.O. Coste ParticleDe M.C. Assis ParticleDe E.A. Marques M.C. Plotkowski (1999) ArticleTitleComparative analysis of three methods to assess viability of mammalian cells in culture Biocell 23 65–72

    Google Scholar 

  • A.G. Doukes T.J. Flotte (1996) ArticleTitlePhysical characteristics and biological effects of laser-induced stress waves Ultrasound Med. Biol. 22 151–64

    Google Scholar 

  • T.A. Drixler M.J. Vogten E.D. Ritchie V.T.J. Van M.F. Gebbink E.E. Voest R.I.H. Borel (2002) ArticleTitleLiver regeneration is an angiogenesis-associated phenomenon Ann. Surg. 236 703–712 Occurrence Handle10.1097/00000658-200212000-00002 Occurrence Handle12454508

    Article  PubMed  Google Scholar 

  • R.I. Freshney (2000) Culture of Animal Cells : A Manual of Basic Technique EditionNumber4 Wiley & Liss Inc New York

    Google Scholar 

  • U. Grossbach (1977) The salivary gland of Chironomus (Diptera): a model system for the study of cell differentiation W. Beermann (Eds) Results and Problems Cell in Differentiation EditionNumberVol. 8 Springer-Verlag Berlin 147–196

    Google Scholar 

  • K.H. Jones J.A. Senft (1985) ArticleTitleAn improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide J. Histochem. Cytochem. 33 77–79 Occurrence Handle1:STN:280:BiqD1c7is1M%3D Occurrence Handle2578146

    CAS  PubMed  Google Scholar 

  • J.P. Kaltenbach M.H. Kaltenbach W.B. Lyons (1958) ArticleTitleNigrosin as a dye for differentiating live and dead ascites cells Exp. Cell. Res. 15 112–117 Occurrence Handle10.1016/0014-4827(58)90067-3 Occurrence Handle1:STN:280:CyaD3czkvFI%3D Occurrence Handle13574164

    Article  CAS  PubMed  Google Scholar 

  • A.W. Krause W.W. Carley W.W. Webb (1984) ArticleTitleFluorescent erythrosin B is preferable to trypan blue as a vital exclusion dye for mammalian cells in monolayer culture J. Histochem. Cytochem. 32 1084–1090 Occurrence Handle1:CAS:528:DyaL2cXmt1KisLY%3D Occurrence Handle6090533

    CAS  PubMed  Google Scholar 

  • D.T. Loo J.R. Rillema (1998) ArticleTitleMeasurement of cell death Methods Cell Biol. 57 251–264 Occurrence Handle1:CAS:528:DyaK1MXmsVan Occurrence Handle9648109

    CAS  PubMed  Google Scholar 

  • T.L. Means D.H. Geroski A. Hadley M.J. Lym H.F. Edelhauser (1995) ArticleTitleViability of human corneal epithelium following optisol-GS storage Arch. Ophthalmol. 113 805–809 Occurrence Handle1:STN:280:ByqA3crnvVA%3D Occurrence Handle7786224

    CAS  PubMed  Google Scholar 

  • T. Mosmann (1983) ArticleTitleRapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays J. Immunol. Methods 65 55–63 Occurrence Handle10.1016/0022-1759(83)90303-4 Occurrence Handle1:STN:280:BiuD1cbgsFE%3D Occurrence Handle6606682

    Article  CAS  PubMed  Google Scholar 

  • B.B. Nath N.N. Godbole (1998) ArticleTitleTechnique for mass rearing of Indian Chironomus species (DipteraNematoceraChironomidae) Studia Dipterologica 5 187–193

    Google Scholar 

  • B.B. Nath S.C. Lakhotia (1989) ArticleTitleHeat shock response and the effect of developmental stage and tissue type on heat shock protein synthesis Genome 32 461–475

    Google Scholar 

  • P. Olinga K. Groen I.H. Hof R.D. Kanter H.J. Koster W.R. Leeman A.A.J.J.L. Rutten K.V. Twillert G.M.N. Groothuis (1997) ArticleTitleComparison of five incubation systems for rat liver slices using functional and viability parameters J. Pharmacol. Toxicol. Methods 38 59–69 Occurrence Handle10.1016/S1056-8719(97)00060-9 Occurrence Handle1:STN:280:DyaK1c%2FmslGmsQ%3D%3D Occurrence Handle9403776

    Article  CAS  PubMed  Google Scholar 

  • T.L. Riss R.A. Moravec (1992) ArticleTitleComparison of MTT, XTT, and a novel tetrazolium compound MTS for in vitro proliferation and chemosensitivity assays Mol. Biol. Cell 3 IssueIDsuppl. 184

    Google Scholar 

  • V. Sorsa (1988) Polytene Chromosomes in Genetic Research Ellis Horwood Ltd Chichester

    Google Scholar 

  • M.G. Stevens S.C. Olsen (1993) ArticleTitleComparative analysis of using MTT and XTT in colorimetric assays for quantitating bovine neutrophil bactericidal activity J. Immunol. Methods 157 225–231 Occurrence Handle10.1016/0022-1759(93)90091-K Occurrence Handle1:CAS:528:DyaK3sXhsVGgt78%3D Occurrence Handle8423367

    Article  CAS  PubMed  Google Scholar 

  • H. Tada O. Shiho K. Kuroshima M. Koyama K. Tsukamoto (1986) ArticleTitleAn improved colorimetric assay for interleukin 2 J. Immunol. Methods 93 157–165 Occurrence Handle10.1016/0022-1759(86)90183-3 Occurrence Handle1:CAS:528:DyaL2sXjtFSjtw%3D%3D Occurrence Handle3490518

    Article  CAS  PubMed  Google Scholar 

  • A.S. Tan M.V. Berridge (2000) ArticleTitleSuperoxide produced by activated neutrophils efficiently reduces the tetrazolium salt, WST-1 to produce a soluble formazan: a simple colorimetric assay for measuring respiratory burst activation and for screening antiinflammatory agents J. Immunol. Methods 238 59–68 Occurrence Handle10.1016/S0022-1759(00)00156-3 Occurrence Handle1:CAS:528:DC%2BD3cXitlOlsrg%3D Occurrence Handle10758236

    Article  CAS  PubMed  Google Scholar 

  • H.N. Traore D. Meyer (2001) ArticleTitleComparing qualitative and quantitative spectroscopic techniques for the detection of the effect of direct iron loading of mammalian cell cultures Methods Cell Sci. 23 175–184 Occurrence Handle10.1023/A:1016313221027 Occurrence Handle1:CAS:528:DC%2BD38XltlemtLY%3D Occurrence Handle12486327

    Article  CAS  PubMed  Google Scholar 

  • H. Yamane K. Konishi H. Iguchi T. Nakagawa S. Shibata M. Takayama K. Nishimura K. Sunami Y. Nakai (1998) ArticleTitleAssessment of hair cell death using the dye extrusion method Acta Otolaryngol. Suppl. 538 7–11 Occurrence Handle1:STN:280:DyaK1M%2FpsFCltA%3D%3D Occurrence Handle9879393

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Zoology, University of Pune, Pune, 411007, India

    B. B. Nath, A. A. Babrekar & B. Parthasarathy

Authors
  1. B. B. Nath
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. A. Babrekar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. B. Parthasarathy
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to B. B. Nath.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nath, B.B., Babrekar, A.A. & Parthasarathy, B. Assessment of Cell Viability in Intact Glandular Tissue in Chironomus ramosus using Dye-exclusion and Colorimetric Assays. Cytotechnology 49, 59–65 (2005). https://doi.org/10.1007/s10616-005-4538-9

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-005-4538-9

Keywords

Search

Navigation