Abstract
The present study introduces the principle of atomic force microscopy (AFM) and reviews our results of human metaphase chromosomes obtained by AFM. AFM imaging of the chromosomes revealed that the chromatid arm was not uniform in structure but had ridges and grooves along its length, which was most prominent in the late metaphase. The arrangement of these ridges and grooves was roughly symmetrical with the counterpart of the paired sister chromatids. AFM imaging of banded chromosomes also showed that the ridges and grooves were related to the G/Q-positive and G/Q-negative bands, respectively. At high magnification, the chromatid was seen to be produced by the compaction of highly twisted chromatin fiber loops, and its compaction tended to be stronger in the ridged regions of the chromosomes than in the grooved regions. Our AFM studies also showed the presence of catenation of chromatin fibers between the ridged portions of the chromatid in the late metaphase. Thus, AFM is useful for obtaining the three-dimensional surface topography not only in ambient conditions but also in physiological liquid conditions, and is expected to be an attractive tool for investigating the structure of chromosomes.
Article PDF
Similar content being viewed by others
References
Bak AL, Zeuthen J, Crick FH (1977) Higher-order structure of human mitotic chromosomes. Proc Nat Acad Sci U S A 74: 1595–1599.
Bath DW (1976) Surface ultrastructure of trypsin-banded chromosomes. Exp Cell Res 98: 262–268.
Bickmore WA, Craig J (1997) Chromosome Bands: Patterns in the Genome. New York: Chapman & Hall.
Binnig G, Quate CF, Gerber C (1986) Atomic force microscope. Phys Rev Lett 56: 930–933.
Caspersson T, Zech L, Johansson C (1970) Differential banding of alkylating fluorochromes in human chromosomes. Exp Cell Res 60: 315–319.
de Grooth BG, Putman CA (1992) High-resolution imaging of chromosome-related structures by atomic force microscopy. J Microsc 168: 239–247.
DuPraw EJ (1965) Macromolecular organization of nuclei and chromosomes: a folded fiber model based on whole-mount electron microscopy. Nature 206: 338–343.
DuPraw EJ (1970) DNA and Chromosomes. New York: Holt, Rinehart and Winston.
Earnshaw WC, Heck MMS (1985) Localization of topoisomerase II in mitotic chromosomes. J Cell Biol 100: 1716–1725.
Engel A, Lyubchenko Y, Müller DJ (1999) Atomic force microscopy: a powerful tool to observe biomolecules at work. Trends Cell Biol 9: 77–80.
Fritzsche W, Schaper A, Jovin TM (1994) Probing chromatin with scanning force microscope. Chromosoma 103: 231–236.
Fritzsche W, Vesenka J, Henderson E (1995) Scanning force microscopy of chromatin. Scanning Microsc 9: 729–739.
Fukushi D, Ushiki T (2005) The structure of C-banded human metaphase chromosomes as observed by atomic force microscopy. Arch Histol Cytol 68: 81–87.
Gall J (1963) Chromosome fibers from an interphase nucleus. Science 139: 120–121.
Harrison CJ, Britch M, Allen TD, Harris R (1981) Scanning electron microscopy of the G-banded human karyotype. Exp Cell Res 134: 141–153.
Harrison CJ, Allen TD, Harris R (1983) Scanning electron microscopy of variations in human metaphase chromosome structure revealed by Giemsa banding. Cytogenet Cell Genet 35: 21–27.
Harrison CJ, Jack EM, Allen TD, Harris R (1985) Light and scanning electron microscopy of the same human metaphase chromosomes. J Cell Sci 77: 143–153.
Holomquist G, Gray M, Porter T, Jordan J (1982) Characterization of Giemsa dark- and light-band DNA. Cell 31: 121–129.
Hörber JK, Miles MJ (2003) Scanning probe evolution in biology. Science 302: 1002–1005.
Hoshi O, Ushiki T (2001) Three-dimensional structure of G-banded human metaphase chromosomes observed by atomic force microscopy. Arch Histol Cytol 64: 475–482.
Hoshi O, Owen R, Miles M, Ushiki T (2004) Imaging of human metaphase chromosomes by atomic force microscopy in liquid. Cytogenet Genome Res 107: 28–31.
Hoshi O, Shigeno M, Ushiki T (2006) Atomic force microscopy of native human metaphase chromosomes in a liquid. Arch Histol Cytol 69: 73–78.
Hoshi O, Hirota T, Kimura E, Komatsubara N, Ushiki T (2007a) Immunocytochemistry for analyzing chromosomes. In: Fukui K, Ushiki T, eds. Chromosome Nanoscience and Technology. Boca Raton, London, New York: CRC Press, pp. 81–91.
Hoshi O, Fukushi D, Ushiki T (2007b) Atomic force microscopy of human chromosomes in relation to their high-order structure. In: Fukui K, Ushiki T, eds. Chromosome Nanoscience and Technology. Boca Raton, London, New York: CRC Press, pp. 105–117.
Inaga S, Tanaka K, Iino A (2002) Three-dimensional helical coiling structures and band patterns of hydrous metaphase chromosomes observed by low vacuum scanning electron microscopy. Arch Histol Cytol 65: 415–423.
Inaga S, Tanaka K, Ushiki T (2007) Transmission and scanning electron microscopy of mammalian metaphase chromosomes. In: Fukui K, Ushiki T, eds. Chromosome Nanoscience and Technology. Boca Raton, London, New York: CRC Press, pp. 93–104.
Iwabuchi S, Muramatsu H, Chiba N et al. (1997) Simultaneous detection of near-field topographic and fluorescence images of human chromosomes via scanning near-field optical/atomic-force microscopy (SNOAM). Nucleic Acids Res 25: 1662–1663.
Kawabata K, Nomura K, Ikeda K et al. (2007) Mechanical approaches to elucidate mechanisms of chromosome condensation at the nano- and micro-level. In: Fukui K, Ushiki T, eds. Chromosome Nanoscience and Technology. Boca Raton, London, New York: CRC Press, pp. 1–13.
Kimura E, Hitomi J, Ushiki T (2002) Scanning neirfield optical/atomic force microscopy of bromodeoxyuridine-incorporated human chromosome. Arch Histol Cytol 65: 435–444.
Kimura E, Hoshi O, Ushiki T (2004) Atomic force microscopy of human metaphase chromosomes after differential staining of sister chromatids. Arch Histol Cytol 67: 171–177.
Kireeva N, Lakonishok M, Kireev I, Hirano T, Belmont AS (2004) Visualization of early chromosome condensation: a hierarchical folding, axial glue model of chromosome structure. J Cell Biol 166: 775–785.
Laemmli UK, Cheng SM, Adolph KW, Paulson JA, Browns JA, Baubach WR (1978) Metaphase chromosome structure and the role of non-histone proteins. Cold Spring Harb Symp Quant Biol 42: 351–360.
Maeshima K, Laemmli UK (2003) A two-step scaffolding model for mitotic chromosome assembly. Dev Cell 4: 467–480.
Makino S (1936) The spiral structure of chromosomes in meiotic division of Podisma (Orthopetera). J Fac Sci Hokkaido Imp Univ, Ser VI 3: 29–40.
Marsden MPF, Laemmli UK (1979) Metaphase chromosome structure: evidence for a radial loop model. Cell 17: 849–868.
Meyer E, Hans JH, Roland B (2004) Scanning Probe Microscopy: The Lab on a Tip. Berlin, Heidelberg: Springer.
Musio A, Mariani T, Frediani C, Ascoli C, Sbrana I (1997) Atomic force microscope imaging of chromosome structure during G-banding treatments. Genome 40: 127–131.
Nomura K, Hoshi O, Fukushi D, Ushiki T, Haga H, Kawabata K (2005) Visualization of elasticity distribution of single human chromosomes by scanning probe microscopy. Jpn J Appl Phys 44: 5421–5424.
Ohnuki Y (1968) Structure of chromosomes. I. Morphological studies of the spiral structure of human somatic chromosomes. Chromosoma 25: 402–428.
Pease RF, Hayes TL (1966) Some biological applications of the scanning electron microscope. In: Ueda R, ed. Electron Microscopy 1966 (Proceedings of 6th International Congress on Electron Microscopy). Tokyo: Marzen, pp. 19–20.
Picco LM, Bozec L, Ulcinas A et al. (2006) Breaking the speed limit with atomic force microscopy. Nanotechnology 18: 1–4.
Putman CAJ, van er Werf KO, de Grooth BG, van Hulst NF, Greve J, Hansma PK (1992) A new imaging mode in Atomic Force Microscopy based on the error signal. SPIE 1639: 198–204.
Rattner JB (1992) Integrating chromosome structure with function. Chromosoma 101: 259–264.
Rattner JB, Lin CC (1985) Radial loops and helical coils coexist in metaphase chromosomes. Cell 42: 291–295.
Sahin FI, Rgun MAE, Tan E, Menevse A (2000) The mechanism of G-banding detected by atomic force microscopy. Scanning 22: 24–27.
Schwarzacher HG, Ruzicka F, Sperling K (1975) Electron microscopy of human banded and prematurely condensed chromosomes. Chromosomes Today 5: 227–234.
Sedat J, Manuelidis L (1978) A direct approach to the structure of eukaryotic chromosomes. Cold Spring Harb Symp Quant Bio 42: 331–350.
Sumner AT (1991) Scanning electron microscopy of mammalian chromosomes from prophase to telophase. Chromosoma 100: 410–418.
Tamayo J, Miles M, Thein A, Soothill P (1999) Selective cleaning of the cell debris in human chromosome preparations studied by scanning force microscopy. J Struct Biol 128: 200–210.
Tamayo J, Miles M (2002) Scanning probe microscopy for chromosomal research. Arch Histol Cytol 65: 369–376.
Tanaka K, Makino R, Iino A (1970) The fine structure of human somatic chromosomes studied by scanning electron microscopy and the replica method. Arch Histol Jpn 32: 203–211.
Thalhammer S, Heckl WM (2003) Atomic force microscopy in cytogenetics. In: Jena BP, Hörber JKH, eds. Force Microscopy: Application in Biology and Medicine. New Jersey: John Wiley & Sons, pp. 249–266.
Thalhammer S, Koehler U, Stark RW, Heckl WM (2001) GTG banding pattern on human metaphase chromosomes revealed by high resolution atomic-force microscopy. J Microsc 202: 46–47.
Ushiki T (2003) Atomic force microscopy for imaging living organism: from DNA to cell motion. In: Fujita H, ed Micromachines as Tools for Nanotechnology. Berlin, Heidelberg: Springer, pp. 121–130.
Ushiki T, Kawabata K (2008) Scanning probe microscopy in biological research. In: Bhushan B, Fuchs H, Tomitori M, eds. Applied Scanning Probe Methods X. Biomimetics and Industrial Applications. Berlin, Heidelberg: Springer, pp. 285–308.
Ushiki T, Hitomi J, Ogura S, Umemoto T, Shigeno M (1996) Atomic force microscopy in histology and cytology. Arch Histol Cytol 59: 421–431.
Ushiki T, Hoshi O, Iwai K, Kimura E, Shigeno M (2002) The structure of human metaphase chromosomes: its histological perspective and new horizons by atomic force microscopy. Arch Histol Cytol 65: 377–390.
White MJD (1940) The heteropycnosis of sex chromosomes and its interpretation in term of spiral structure. J Genet 40: 67–82.
Wray W, Stubblefield E (1970) A new method for the rapid isolation of chromosomes, mitotic apparatus, or nuclei from mammalian fibroblasts at near neutral pH. Exp Cell Res 59: 469–478.
Yoshino T, Sugiyama S, Hagiwara S, Ushiki T, Ohtani T (2002) Simultaneous collection of topographic and fluorescent images of barley chromosomes by scanning near-field optical/atomic force microscopy. J Electron Microsc 51: 199–203.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ushiki, T., Hoshi, O. Atomic force microscopy for imaging human metaphase chromosomes. Chromosome Res 16, 383–396 (2008). https://doi.org/10.1007/s10577-008-1241-7
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10577-008-1241-7