In situ hybridization to chromosomes stabilized in gel microdrops.
Conventional chromosome in situ hybridization procedures rely on fixation to glass slides followed by microscopic evaluation. This report describes the development of a microdrop in situ hybridization to chromosomes in suspension. Chromosomes encapsulated in gel microdrops (GMDs) composed of an agarose matrix withstood stringent hybridization and denaturation conditions. Because of the increased stability, hybridization to encapsulated chromosomes was detected by flow cytometry as well as conventional microscopy. Thus, the MISH method offers a means for chromosome hybridization without slides and may enable identification and isolation of chromosome using hybridization rather than nucleic acid binding dyes.
Nguyen
B T
BT
One Cell Systems, Inc., Cambridge, Massachusetts 02139, USA.
Lazzari
K
K
Abebe
J
J
Mac
I
I
Lin
J B
JB
Chang
A
A
Wydner
K L
KL
Lawrence
J B
JB
Cram
L S
LS
Weier
H U
HU
D016475:3T3 Cells; D000818:Animals; D002875:Chromosomes / Q000502:physiology / Q000648:ultrastructure*; D002877:Chromosomes, Human / Q000502:physiology / Q000648:ultrastructure*; D005434:Flow Cytometry / Q000379:methods; D005782:Gels; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
D005782:Gels