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İkili İskelet Boyamaları

Double Skeletal Staining

Journal Name:

  • Uludağ Üniversitesi Tıp Fakültesi Dergisi

Publication Year:

  • 2005

Key Words:

Keywords (Original Language):

Author NameUniversity of Author
Abstract (2. Language): 
Because of its trustable results, double skeletal staining is the most widely used method in experimental teratological studies and should be known very well by the researchers who work teratology. This review aims to form a source for the teratology researchers by summarizing the developments about the skeletal staining from the first day to today. The first important development of skeletal staining has been the staining of bone with alizarin red S. After that, because the skeleton of the foetuses of laboratory animals at term contains a considerable amount of cartilage, it has been realized that staining of the bone alone is insufficient and search for a dye, which will stain the cartilage properly, has been started. For this aim, methylen blue, toludin blue, methyl green and rezorsin fuksin has been tested, but the satisfactory result were obtained with alcian blue. After the publication of double skeletal staining methods in which these two dyes were used; studies which aim to increase the quality of the staining, to decrease the duration of the staining, to simplify the staining method, to use the skeletal staining with other methods and to describe methods which can be used in different species has been done.
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Abstract (Original Language): 
İkili iskelet boyaması, sonuçlarının güvenilir olması nedeniyle; deneysel teratolojik çalışmalarda en sık kullanılan yöntemdir ve bu nedenle de teratoloji çalışan araştırmacılar tarafından iyi bilinmesi son derece önemlidir. Bu derlemenin amacı; iskelet boyamasında, ilk uygulanmaya başlandığı tarihlerden günümüze kadar gerçekleşen başlıca gelişmeleri özetleyerek araştırmacılara kaynak oluşturmaktır. İskelet boyamalarında ilk önemli gelişme kemiğin alizarin red S ile boyanmasıdır. Ardından, termdeki deney hayvanı fetuslarının iskeletinin önemli bir bölümünün kıkırdaktan oluşması nedeniyle, sadece kemiğin boyanmasının yeterli olmayacağı düşünülerek kıkırdak bölümleri boyayacak uygun ajanlar aranmaya başlanmıştır. Bu amaçla, methylen blue, toludin blue, methyl green ve rezorsin fuksin denenmiş, ancak tatmin edici sonuçların alındığı boya alcian blue olmuştur. Bu iki boyanın beraber kullanıldığı ikili iskelet boyaması yöntemlerinin yayınlanmasının ardından da; boyanma kalitesini arttırmayı, işlemin süresini kısaltmayı, erişkin deney hayvanlarında kullanılabilecek metotlar tanımlamayı, boyama işlemini daha basit ve daha pratik hale getirmeyi, iskelet boyamalarını farklı amaçlara sahip diğer teknikler ile bir arada kullanmayı ve farklı türlerin fetuslarında kullanılabilecek metotlar geliştirmeyi amaçlayan çalışmalar yapılmıştır.
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  • 2
119-126
  • Turkish

REFERENCES

References: 

1. Lee M, Leichter J. Skeletal development in fetuses of rats
consuming alcohol during gestation. Growth. 1983; 47: 254–
62.
2. Alles AJ, Sulik KK. Retinoic acid induced limb reduction
defects: Perturbation of zones of programmed cell death as a
pathogenetic mechanism. Teratology 1989; 40: 163–71.
3. Bannigan JG, Cottell DC, Morris A. Study of mechanism of
BUdR–induced cleft palate in the mouse. Teratology 1990;
42: 79–89.
4. Tanigawa K, Kawaguchi M, Tanaka O, Kato Y. Long–term
effect of maternal insulin–induced hypoglycemia during organogenesis. Diabetes 1991; 40: 1115–21.
5. Ehlers K, Sturge H, Merker H–J, Nau H. The valproic acid
metabolite E–2–n–Propyl–2–Pentenoic Acid does not induce
spina bifida in the mouse. Dev Pharmacol Ther 1992; 19:
196–204.
6. Savontaus M, Metsaranta M, Vuorio E. Retarded skeletal
development in transgenic mice with a Type II collagen mutation. Am J Pathol 1996; 149: 2169–82.
7. Migliaccio S, Newbold RR, Bullock BC, Jefferson WJ, Sutton
FG, Mclachlan JA, Korach KS. Alterations of maternal levels
during gestation affect the skeleton of female offspring. Endocrinology 1996; 137: 2118–25.
8. Savontaus M, Metsaranta M, Vuorio E. Mutation in type II
collagen gene disturbs spinal development and gene expression patterns in transgenic Del1 mice. Lab Invest 1997; 77:
591–600. İkili İskelet Boyamaları
125
9. Maddox BK, Garofalo S, Smith C, Keene DR, Horton WA.
Skeletal development in transgenic mice expressing a mutation at Gly574Ser of Type II collagen. Dev Dynam 1997; 208:
170–7.
10. Bareggi R, Grill V, Sandrucci MA, Baldini G, De Pol A,
Forabosco A, Narducci P. Developmental pathways of vertebral centra and neural arches in human embryos and fetuses.
Anat. Embryol 1993; 187: 139–44.
11. Bareggi R, Grill V, Narducci P, Forabosco A. A quantitative
study on the spatial and temporal ossification patterns of vertebral centra and neural arches and their relationship to the fetal age. Ann. Anat 1994; 176: 311–7.
12. Bareggi R, Grill V, Zweyer M, Sandrucci MA, Narducci P,
Forabosco A. The growth of long bones in human embryological and fetal upper limbs and its relationship to other developmental patterns. Anat. Embryol. 1994; 189: 19–24.
13. Tomo S, Ogita M, Tomo I. Development of mandibular
cartilages in the rat. Anat Rec. 1997; 249: 233–9.
14. Danielson M, Kihlstrom I. Calcification of the rabbit fetal
skeleton. Growth. 1986; 50: 378–84.
15. Menegola E, Broccia ML, Giavini E. Atlas of rat fetal skeleton double stained for bone and cartilage. Teratology 2001;
64: 125–33.
16. Hill MF, Zdan R. Staining of developing skeleton with murexide. Acta anat. 1973; 84: 509–15.
17. Dawson AB. A note on the staining of the skeleton of cleared
specimens with Alizarin red–S. Stain Technol. 1926; 1: 123–
4.
18. Lipman H. Staining the skeleton of cleared embryos with
alizarin red–S. Stain Technol. 1935; 10: 61–3.
19. Mitala JJ, Boardman JP, Carrano RA, Iuliucci JD. Novel
accessory skull bone in fetal rats after exposure to Aspirin.
Teratology. 1984; 30: 95–8.
20. Virtanen P, Lassila V. Alizarin red–S stained bone and cartilage in calcium deficiency provoked by experimental liver injury in rats. Acta Anat. 1986; 125: 6–9.
21. Curle DC, Ray M, Persaud TVN. In vivo evaluation of teratogenesis and cytogenetic changes following methylmercuric
chloride treatment. Anat Rec. 1987; 219: 286–95.
22. Cassella JP, Pereira R, Khillan DJ, Prockop DJ, Garrington N,
Ali SY. An ultrastuructural microanalytical and spectroscopic
study of bone from a transgenic mouse with a COL1.A1 pro–
alpha–1 mutation. Bone. 1994; 15: 611–9.
23. Ojeda JL, Barbosa E, Bosque PG. Selective skeletal staining
in whole chicken embryos; A rapid alcian blue technique.
Stain Technol. 1970; 45: 137–9.
24. Love AM, Vickers TH. Durable staining of cartilage in foetal
rat skeleton by methylene blue. Stain Technol. 1972; 47: 7–
11.
25. Kimmel CA, Laborde JB, Trammell CT. Evaluation of cartilage and bone formation in fetal skeletons following prenatal
insult reveals abnormalities not apparent in alizarin stained
specimens. Teratology. 25: 54A–55A.
26. Williams TW. Alizarin red–S and toluidine blue for differentiating adult or embryonic bone and cartilage. Stain Technol.
1941; 16: 23–5.
27. Burdi A. Toluidine blue Alizarin red–S staining of cartilage
and bone in whole–mount skeletons in vitro. Stain Technol.
1965; 40: 45–8.
28. Burdi AR, Flecker K. Differential staining of cartilage and
bone in the intact chick embryonic skeleton in Vitro. Stain
Technol. 1968; 43: 47–8.
29. Inouye M. Differential staining of cartilage and bone in fetal
mouse skeleton by Alcian blue and Alizarin red S. Cong
Anom. 1976; 16: 171–3.
30. Wassersug RJ. A procedure for differential staining of cartilage and bone in whole formalin fixed vertebrates. Stain
Technol. 1976; 51: 131–4.
31. Dingerkus G, Uhler LD. Enzyme clearing of alcian blue
stained whole small vertebrates for demonstration of cartilage.
Stain Technol. 1977; 52: 229–32.
32. Mc Cann JA. Methyl green as a cartilage stain; human embryos. Stain Technol. 1971; 46: 263–5.
33. Kimmel CA, Trammell C. A rapid procedure for routine
double staining of cartilage and bone in fetal and adult animals. Stain Technol. 1981; 56: 271–3.
34. Rousseaux CG. Automated differential staining for cartilage
and bone in whole mount preparations of vertebrates. Stain
Technol. 1985; 60: 295–7.
35. Miller DM, Tarpley J. An automated double staining procedure for bone and cartilage. Biotech Histochem. 1996; 71: 79–
83.
36. Kelly WL, Bryden MM. A modified stain for cartilage and
bone in whole mount preparations of mammalian fetuses and
small vertebrates. Stain Technol. 1983; 58: 131–4.
37. Whitaker J, Dix KM. Double staining technique for rat foetus
skeletons in teratological studies. Lab Anim. 1979; 13: 309–
10.
38. Trueman D, Jackson SW, Trueman B. An automated technique for double staining rat and rabbit fetal skeletal specimens to differentiate bone and cartilage. Biotech Histochem.
1999; 74: 98–104.
39. Tarpley JE. Adult rodent double skeleton stain. Biotech
Histochem. 1998; 74: 116–8.
40. Park E–H, Kim DS. A procedure for staining cartilage and
bone of whole vertebrate larvae while rendering all other tissues transparent. Stain Technol. 1984; 59: 269–72.
41. Webb GN, Byrd RA. Simultaneous differential staining of
cartilage and bone in rodent fetuses: an Alcian blue and Alizarin red S procedure without glacial acetic acid. Biotech Histochem. 1994; 69: 181–5.
42. Yamada T. Selective staining methods for cartilage of rat fetal
specimens previously treated with Alizarin red S. Teratology.
1991; 43: 615–9.
43. Yamada T, Kurihara A, Nishiyama T, Uchida H. Application
of the bromophenol blue staining method to rat fetal cartilage
previously stained with Alizarin red S. Exp Anim. 1993; 42:
457–61.
44. Hood RC, Neill WM. A modification of alizarin red–S technic
for demonstrating bone formation. Stain Technol. 1948; 23:
209–18.
45. Sedra S. Decreasing the time required for making an alizarin
skeleton preparation. Stain Technol. 1950; 25: 223–4.
46. Taylor WR. An enzyme method of clearing and staining small
vertebrates. Proc. U. S. Nat. Mus. 1967; 122: 1–17.
47. Taylor WR. Outline of a method of clearing tissues with
pancreatic enzymes and staining bones of small vertebrates.
Turtox News. 1967; 45: 308–9.
48. Gosztonyi AE. The use of enzyme–based laundry ″ presoaks ″
for clearing small vertebrates for alizarin red staining of bony
tissues. Stain Technol. 1984; 59: 305–7.
49. Chappard D, Alexandre C, Palle S, Rıffat G. Permanent
preservation of whole alizarin red S skeletons by clearing and
embedding in polyester resins. Stain Technol. 1985; 61: 145–
9.
50. Jensh RP, Brent RL: Rapid schedules for KOH clearing and
alizarin red S staining of fetal rat bone. Stain Technol. 1966;
41: 179–83.
51. Carlson BM, Simandl BK, Stocker KM, Connelly TG, Fallon
JF: A method for combined gross skeletal staining and feulgen F.B. Sunay
126
staining of embryonic chick tissues. Stain Technol. 1986; 61:
27–31.
52. True RM. Staining of embryonic and small mammalian skeletal systems. Stain Technol. 1947; 22: 107–8.
53. Crary DD. Modified benzyl alcohol clearing of alizarin
stained specimens without loss of flexibility. Stain Technol.
1962; 37: 124–5.
54. Selby PB. A rapid method for preparing high quality alizarin
stained skeletons of adult mice. Stain Technol. 1987; 62: 143–
6.
55. Petrere JA, Schardeın JL. Microwave fixation of fetal specimens. Stain Technol. 1977; 52: 113–4.
56. Kahveci Z, Çavuşoğlu İ, Sırmalı ŞA. Microwave fixation of
whole fetal specimens. Biotech Histochem. 1997; 72: 144–7.
57. Sunay FB. İkili iskelet boyamalarında mikrodalga ışınımının
kullanılması (Uzmanlık Tezi). Bursa: Uludağ Üniversitesi;
2000.
58. Boardman JP, Mitala JJ, Carrano RA, Iuliucci JD: Cartilage–
staining technique for the examination of unskinned fetal rat
specimens previously processed with Alizarin red S. Teratology. 1984; 30: 383–4.
59. Dudzinski KM, Neff NA. A technique for the combination of
clearing, staining, and injecting small mammals. Stain Technol. 1990; 65: 113–8.
60. Dingerkus G. The use of various alcohols for alcian blue in
toto staining of cartilage. Stain Technol. 1981; 56: 128–9.
61. Peltzer MA, Schardeın JL: A convenient method for processing fetuses for skeletal staining. Stain Technol. 1966; 41:
1420–1.
62. Burdan F, Rozylo–Kalinowska I, Katarzyna Rozylo T, Chahoud I: A new radip procedure for routine teratological use in
bone ossification assessment: a supplement for staining methods. Teratology. 2002; 66: 315–25.
63. Rozylo–Kalinowska I; Michalska A, Burdan F: Optimization
of analysis of skeletal ossification of laboratory animals by
means of digital radiography software options. Ann Univ
Mariae Curie Sklodowska. 2003; 58:95-100.

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