In situ hybridization at the ultrastructural level: localization of cytomegalovirus DNA using digoxigenin labelled probes.
We describe a procedure for detecting and localizing cytomegalovirus DNA sequences based on in situ hybridization at the ultrastructural level. A digoxigenin-labelled probe, identified with an anti-digoxigenin colloidal gold-labelled antiserum, was employed on infected cells embedded in a new acrylic resin (Bioacryl). The silver enhancement method on the same specimen was used to more easily reveal the reaction also at low magnification. The immunolocalization was characterized by high specificity with virtually no background staining and a good maintenance of submicroscopic cell features.
Cenacchi
G
G
Institute of Clinical Electron Microscopy, S. Orsola Hospital, University of Bologna, Italy.
Musiani
M
M
Gentilomi
G
G
Righi
S
S
Zerbini
M
M
Chandler
J G
JG
Scala
C
C
La Placa
M
M
Martinelli
G N
GN
D000180:Acrylic Resins; D000918:Antibody Specificity; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics*; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004076:Digoxigenin; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology / Q000382:microbiology; D006801:Humans; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008168:Lung / Q000737:chemistry / Q000166:cytology / Q000382:microbiology; D008854:Microscopy, Electron; D012834:Silver
D000180:Acrylic Resins; D015342:DNA Probes; D004279:DNA, Viral; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D012834:Silver; D004076:Digoxigenin