Detection of genetic alterations by immunoFISH analysis of whole cells extracted from routine biopsy material.
The detection of genetic abnormalities (eg, translocations, amplifications) in paraffin-embedded samples by the fluorescence in situ hybridization (FISH) technique is usually performed on tissue sections. FISH analysis of nuclei extracted from paraffin-embedded samples is also possible, but the technique is not widely used, principally because of the extra labor involved and the loss of information on tissue architecture. In this article, we report that nuclei extracted from paraffin-embedded tissue often retain at least part of the surrounding cytoplasm. Consequently, immunocytochemical labeling for a range of cellular markers (eg, of lineage or proliferation) can be performed in combination with FISH labeling, allowing specific cell populations to be analyzed for genetic abnormalities. These cell preparations are largely free of the problems associated with tissue sections (eg, truncation artifact, signals in different focal planes) so that interpretation is easy and numerical chromosomal abnormalities are readily assessed. Cells isolated from paraffin sections can be stored in suspension so that arrays can be created as and when needed from a range of neoplasms for investigation by the immunoFISH technique (for example, for studying a new genetic abnormality). This procedure represents a novel methodology, which in some settings offers clear advantages over analysis of tissue sections.
Mattsson
Göran
G
Haematology Department, Level 4, John Radcliffe Hospital, Oxford OX3 9DU, UK.
Tan
Soo Yong
SY
Ferguson
David J P
DJ
Erber
Wendy
W
Turner
Susan H
SH
Marafioti
Teresa
T
Mason
David Y
DY
D001706:Biopsy; D002457:Cell Extracts; D002469:Cell Separation; D002869:Chromosome Aberrations; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D009369:Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D014066:Palatine Tonsil / Q000473:pathology / Q000648:ultrastructure; D016612:Paraffin Embedding; D016707:Tissue Fixation
D002457:Cell Extracts
Br J Haematol. 1996 Mar;92(3):673-80
8616034
Br J Haematol. 1995 Jul;90(3):701-6
7647013
Hum Pathol. 1999 Mar;30(3):324-30
10088552
Diagn Mol Pathol. 2004 Dec;13(4):197-206
15538109
J Pathol. 2005 Feb;205(3):302-10
15682432
Eur J Histochem. 2005 Jan-Mar;49(1):53-8
15823795
Leukemia. 2005 May;19(5):856-61
15744341
J Pathol. 2005 Jun;206(2):233-41
15822070
J Clin Pathol. 2005 Dec;58(12):1336-8
16311361
Mod Pathol. 2006 Jan;19(1):1-8
16258512
Blood. 2006 Jan 1;107(1):213-21
16160011
J Mol Diagn. 2006 May;8(2):141-51
16645199
Lab Invest. 2006 Jun;86(6):547-56
16607381
Haematologica. 2006 Jun;91(6):772-80
16769579
Appl Immunohistochem Mol Morphol. 2006 Jun;14(2):220-4
16785794
J Pathol. 2006 Aug;209(4):454-63
16739114
J Mol Diagn. 2006 Sep;8(4):459-65
16931586
J Cancer Res Clin Oncol. 2007 Feb;133(2):117-23
16977458
Pediatr Dev Pathol. 2000 May-Jun;3(3):249-55
10742412
Virchows Arch. 2000 Mar;436(3):271-5
10782886
J Pathol. 2000 Aug;191(4):452-61
10918222
Br J Haematol. 2000 Sep;110(4):856-62
11054068
Mol Pathol. 2002 Apr;55(2):121-4
11950961
Br J Haematol. 2002 May;117(2):322-32
11972514
Am J Pathol. 2002 Jun;160(6):1967-72
12057901
J Pathol. 2002 Oct;198(2):163-70
12237875
Br J Haematol. 2003 Apr;121(1):49-56
12670331
Clin Cancer Res. 2003 Jun;9(6):2133-9
12796378
Mod Pathol. 2003 Jul;16(7):708-15
12861068
J Mol Diagn. 2003 Aug;5(3):168-75
12876207
Am J Clin Pathol. 2003 Sep;120(3):327-34
14502796
Blood. 2004 Jan 1;103(1):188-93
12881301
Leukemia. 2004 Feb;18(2):348-53
14671641
Br J Haematol. 2004 Feb;124(4):519-33
14984504
Diagn Mol Pathol. 2004 Jun;13(2):97-104
15167011
Hum Pathol. 2004 Jun;35(6):752-8
15188143
N Engl J Med. 2004 Jul 15;351(3):250-9
15254283
Cancer Genet Cytogenet. 1998 Jul 15;104(2):146-52
9666809