title: Selective cleaning of the cell debris in human chromosome preparations studied by scanning force microscopy.
abstract: The chromosome structure is one of most challenging biological structures to be discovered. Most evidence about the structure comes from optical microscopy. Scanning force microscopy (SFM) can achieve molecular resolution and allows imaging in liquids. However, little information about the chromosome structure has been revealed by SFM. In this work, a mild enzymatic treatment is applied to the chromosomes to remove selectively the RNA and proteins coming from the cell. The resulting SFM images indicate that a protein film with embedded RNA molecules covers chromosomes in standard cytogenetic preparations. The thickness of the protein layer is 15-35 nm and the RNA adheres preferentially to the chromosome surface. The cell material film results in a quite smooth chromosome surface without evidence of any structural detail. After treatment, the chromosome was cleaned from cell residues and individual chromatin fibers at the surface were resolved. Furthermore, insights about the higher order structure of the chromosome can be inferred.
authors.affiliation: H. H. Wills Physics Laboratory, Fetal Medicine Research Unit, St. Michael's Hospital, University of Bristol, Tyndall Avenue, Bristol, BS8 1TL, United Kingdom. phjt@phys.bris.ac.uk
authors.forename: J
authors.initials: J
authors.lastname: Tamayo
authors.forename: M
authors.initials: M
authors.lastname: Miles
authors.forename: A
authors.initials: A
authors.lastname: Thein
authors.forename: P
authors.initials: P
authors.lastname: Soothill
mesh_terms: D002843:Chromatin / Q000648:ultrastructure; D002877:Chromosomes, Human / Q000648:ultrastructure*; D006801:Humans; D007962:Leukocytes; D018625:Microscopy, Atomic Force / Q000379:methods*; D010434:Pepsin A; D012260:Ribonucleases
chemical_list: D002843:Chromatin; D012260:Ribonucleases; D010434:Pepsin A