US 12,253,468 B2
Methods for fluorescence imaging microscopy and nano-fish
John A. Stamatoyannopoulos, Seattle, WA (US); Shreeram Akilesh, Seattle, WA (US); and Vivek Nandakumar, Seattle, WA (US)
Assigned to Altius Institute for Biomedical Sciences, Seattle, WA (US)
Filed by Altius Institute for Biomedical Sciences, Seattle, WA (US)
Filed on May 4, 2022, as Appl. No. 17/736,684.
Application 17/736,684 is a continuation of application No. 16/319,157, granted, now 11,353,400, previously published as PCT/US2017/042946, filed on Jul. 19, 2017.
Claims priority of provisional application 62/364,245, filed on Jul. 19, 2016.
Prior Publication US 2022/0397526 A1, Dec. 15, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 21/64 (2006.01); A01N 1/02 (2006.01); C12N 5/078 (2010.01); C12N 5/09 (2010.01); C12P 19/34 (2006.01); C12Q 1/6837 (2018.01); C12Q 1/6841 (2018.01); C40B 40/06 (2006.01); G01N 1/30 (2006.01); G01N 21/78 (2006.01); G01N 33/569 (2006.01); G01N 33/574 (2006.01); G01N 33/58 (2006.01); C40B 30/04 (2006.01); G01N 15/00 (2006.01)
CPC G01N 21/64 (2013.01) [A01N 1/0221 (2013.01); C12N 5/0634 (2013.01); C12N 5/0694 (2013.01); C12P 19/34 (2013.01); C12Q 1/6837 (2013.01); C12Q 1/6841 (2013.01); C40B 40/06 (2013.01); G01N 1/30 (2013.01); G01N 21/6458 (2013.01); G01N 21/78 (2013.01); G01N 33/56966 (2013.01); G01N 33/574 (2013.01); G01N 33/582 (2013.01); C40B 30/04 (2013.01); G01N 2015/0038 (2013.01)] 14 Claims
 
1. A high-throughput method for detecting presence of a 800 nucleotides to 2.1 kb long target nucleic acid sequence in a genome of mammalian cells with high resolution, the method comprising:
contacting the mammalian cells with a plurality of probes comprising a detectable label and a probe sequence that binds to a portion of the target nucleic acid sequence; and
detecting a presence of the detectable label localized to a region in the genome in the nucleus of the mammalian cells, wherein the presence of the detectable label indicates the presence of the target nucleic acid sequence,
wherein the target nucleic acid is an endogenous nucleic acid sequence present in the genome of the cells, an exogenous nucleic acid sequence integrated into the genome of the cells, or a combination thereof,
wherein the probe sequence is between 30 and 50 nucleotides in length,
wherein the plurality of probes comprises at least 30 probes to less than 50 probes, and
wherein high resolution comprises delineation of 800 nucleotides to 2.1 kb long target sequences separated by a 4 kb intervening sequence on a chromosome.