PMID- 14673657
OWN - NLM
STAT- MEDLINE
DCOM- 20040914
LR  - 20181113
IS  - 0948-6143 (Print)
IS  - 0948-6143 (Linking)
VI  - 121
IP  - 1
DP  - 2004 Jan
TI  - Dual Y chromosome painting and in situ cell-specific immunofluorescence staining 
      in lung tissue: an improved method of identifying donor marrow cells in lung 
      following bone marrow transplantation.
PG  - 73-9
AB  - Several recent studies have demonstrated localization of donor bone 
      marrow-derived cells in recipient lungs following transplantation from male to 
      female mice or patients. Donor cells are identified by detection of the Y 
      chromosome by fluorescence in situ hybridization (FISH). However, protein 
      digestion pretreatments usually required for tissue FISH significantly limit the 
      ability to detect cell type-specific markers by immunohistochemistry. We have 
      used an alternative protein digest approach that entails heating the slides in 10 
      mM sodium citrate rather than utilizing a protease digestion. This approach 
      preserves cell proteins following FISH, and allows lung tissue to remain intact 
      for subsequent detection of cell-specific markers by immunohistochemistry. We 
      have examined this technique in mouse lungs using a Y chromosome paint and three 
      cell-specific markers, a pan-cytokeratin for epithelial cells, PECAM-1 for 
      endothelial cells, and CD45 for leukocytes. Excellent visualization of both the Y 
      chromosome and cell-specific surface protein markers was obtained on a single 
      slide. This approach will significantly enhance the ability to detect and 
      identify donor bone marrow cells in recipient mouse lungs following male to 
      female transplantation.
FAU - Trotman, Winifred
AU  - Trotman W
AD  - Department of Pathology, University of Vermont College of Medicine, Burlington, 
      Vermont 05405, USA.
FAU - Beckett, Travis
AU  - Beckett T
FAU - Goncz, Kaarin K
AU  - Goncz KK
FAU - Beatty, Barbara G
AU  - Beatty BG
FAU - Weiss, Daniel J
AU  - Weiss DJ
LA  - eng
PT  - Journal Article
DEP - 20031212
PL  - Germany
TA  - Histochem Cell Biol
JT  - Histochemistry and cell biology
JID - 9506663
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/*cytology
MH  - *Bone Marrow Transplantation
MH  - Chromosome Painting
MH  - Female
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - In Vitro Techniques
MH  - Lung/*chemistry/*cytology/ultrastructure
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - *Y Chromosome/ultrastructure
EDAT- 2003/12/16 05:00
MHDA- 2004/09/15 05:00
CRDT- 2003/12/16 05:00
PHST- 2003/10/29 00:00 [accepted]
PHST- 2003/12/16 05:00 [pubmed]
PHST- 2004/09/15 05:00 [medline]
PHST- 2003/12/16 05:00 [entrez]
AID - 10.1007/s00418-003-0598-0 [doi]
PST - ppublish
SO  - Histochem Cell Biol. 2004 Jan;121(1):73-9. doi: 10.1007/s00418-003-0598-0. Epub 
      2003 Dec 12.