Forensic applications of laser capture microdissection: use in DNA-based parentage testing and platform validation.
OBJECTIVE
To report on the successful use of Laser Capture Microdissection (LCM) as a tool for isolation of human chorionic villi from admixed maternal tissue. Subsequent DNA isolation for forensic short tandem repeat (STR) analysis for parentage testing was performed in two cases of alleged sexual assault of female victims. We also performed validation of the LCM instrument platform, using archival formalin-fixed human fetal products of conception (POC), for which microdissection was utilized to separate maternal (decidua) and fetal (chorionic villus) components.


METHODS
To isolate DNA from placental chorionic villi admixed with maternal decidua recovered after spontaneous or therapeutic abortion, LCM was used to separate fetal from maternal cells. In contrast to the relatively crude conventional microdissection performed using a narrow pipette, needle, or scalpel blade, LCM allows cell- or tissue-specific isolation of placental chorionic villi from archival paraffin-embedded tissue sections, leaving the maternal tissue intact.


RESULTS
After polymerase chain reaction (PCR) amplification of villi after LCM of 9-15 STR loci, the quantity and quality of DNA yielded from fetal cells isolated by LCM was sufficient for PCR analysis and successful forensic parentage testing. The validation data obtained on two sets of formalin-fixed archival POC tissues from anonymous donors demonstrated the encouraging reproducibility of these protocols and procedures.


CONCLUSIONS
We demonstrated the reliability and utility of LCM for forensic applications when high specificity of a particular analyzed cell population or tissue is required. Care must be taken during routine pathology procedures to avoid contamination of tissues with admixture of extraneous DNA.
Budimlija
Zoran M
ZM
Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, 75 Francis Street, Boston, MA 02115, USA.
Lechpammer
Mirna
M
Popiolek
Dorota
D
Fogt
Franz
F
Prinz
Mechthild
M
Bieber
Frederick R
FR
D000293:Adolescent; D002824:Chorionic Villi; D004247:DNA / Q000235:genetics*; D005260:Female; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D010334:Paternity; D011247:Pregnancy
D004247:DNA