title: Performance of computer vision in vivo flow cytometry with low fluorescence contrast.
abstract: Detection and enumeration of circulating cells in the bloodstream of small animals are important in many areas of preclinical biomedical research, including cancer metastasis, immunology, and reproductive medicine. Optical in vivo flow cytometry (IVFC) represents a class of technologies that allow noninvasive and continuous enumeration of circulating cells without drawing blood samples. We recently developed a technique termed computer vision in vivo flow cytometry (CV-IVFC) that uses a high-sensitivity fluorescence camera and an automated computer vision algorithm to interrogate relatively large circulating blood volumes in the ear of a mouse. We detected circulating cells at concentrations as low as 20 cells/mL. In the present work, we characterized the performance of CV-IVFC with low-contrast imaging conditions with (1) weak cell fluorescent labeling using cell-simulating fluorescent microspheres with varying brightness and (2) high background tissue autofluorescence by varying autofluorescence properties of optical phantoms. Our analysis indicates that CV-IVFC can robustly track and enumerate circulating cells with at least 50% sensitivity even in conditions with two orders of magnitude degraded contrast than our previous in vivo work. These results support the significant potential utility of CV-IVFC in a wide range of in vivo biological models.
authors.forename: Stacey
authors.initials: S
authors.lastname: Markovic
authors.forename: Siyuan
authors.initials: S
authors.lastname: Li
authors.forename: Mark
authors.initials: M
authors.lastname: Niedre
mesh_terms: D000465:Algorithms; D000818:Animals; D001772:Blood Cell Count / Q000295:instrumentation / Q000379:methods*; D001773:Blood Cells; D003201:Computers; D003287:Contrast Media; D054644:Ear Auricle / Q000098:blood supply; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008863:Microspheres; D019047:Phantoms, Imaging; D012680:Sensitivity and Specificity
chemical_list: D003287:Contrast Media
references.citation: Eur Radiol. 2008 Oct;18(10):2021-32
references.pmid: 18506449
references.citation: Clin Cancer Res. 2008 Oct 1;14(19):6302-9
references.pmid: 18829513
references.citation: J Biomed Opt. 2005 Jul-Aug;10(4):41202
references.pmid: 16178626
references.citation: Opt Lett. 2007 Dec 1;32(23):3450-2
references.pmid: 18059963
references.citation: Sci Rep. 2013;3:1259
references.pmid: 23405273
references.citation: Nat Methods. 2007 Sep;4(9):741-6
references.pmid: 17721542
references.citation: J Clin Oncol. 2008 Jul 1;26(19):3213-21
references.pmid: 18591556
references.citation: Phys Med Biol. 2012 Jul 21;57(14):4627-41
references.pmid: 22750660
references.citation: Nat Med. 2010 Jun;16(6):718-22
references.pmid: 20495571
references.citation: Methods. 2010 Apr;50(4):289-97
references.pmid: 20116432
references.citation: Cytometry A. 2011 Oct;79(10):848-54
references.pmid: 21948732
references.citation: Cytometry A. 2011 Oct;79(10):775-88
references.pmid: 21796772
references.citation: Cancer Res. 2004 Aug 1;64(15):5044-7
references.pmid: 15289300
references.citation: J Biomed Opt. 2012 Mar;17(3):037001
references.pmid: 22502573
references.citation: Nat Methods. 2010 Oct;7(10):827-9
references.pmid: 20818379
references.citation: J Biomed Opt. 2008 Jul-Aug;13(4):044008
references.pmid: 19021336
references.citation: J Biophotonics. 2009 Sep;2(8-9):457-8
references.pmid: 19743439
references.citation: Opt Lett. 2005 Mar 15;30(6):628-30
references.pmid: 15791998
references.citation: Cancers (Basel). 2013 Dec 10;5(4):1691-738
references.pmid: 24335964
references.citation: Opt Lett. 2004 Jan 1;29(1):77-9
references.pmid: 14719666
references.citation: Methods. 2012 Jul;57(3):280-96
references.pmid: 22749928
references.citation: J Biomed Opt. 2013 Jul;18(7):077002
references.pmid: 23831714
references.citation: Prog Mol Biol Transl Sci. 2010;95:95-112
references.pmid: 21075330
references.citation: Cancer Res. 2009 Oct 15;69(20):7926-34
references.pmid: 19826056
references.citation: N Engl J Med. 2004 Aug 19;351(8):781-91
references.pmid: 15317891
references.citation: Cytometry A. 2011 Oct;79(10):737-45
references.pmid: 21915991
references.citation: Nature. 2007 Dec 20;450(7173):1235-9
references.pmid: 18097410
references.citation: J Assist Reprod Genet. 2003 Feb;20(2):58-62
references.pmid: 12688589
references.citation: Lancet Oncol. 2004 Feb;5(2):79-88
references.pmid: 14761811
references.citation: Cytometry A. 2013 Dec;83(12):1113-23
references.pmid: 24273157
references.citation: Opt Lett. 2013 Jul 1;38(13):2357-9
references.pmid: 23811927
references.citation: J Biomed Opt. 2011 Jan-Feb;16(1):011006
references.pmid: 21280893
references.citation: Nat Rev Cancer. 2005 Aug;5(8):591-602
references.pmid: 16056258
references.citation: Cytometry A. 2011 Oct;79(10):758-65
references.pmid: 21905206
references.citation: Science. 2006 Apr 14;312(5771):217-24
references.pmid: 16614209
references.citation: Ann Intern Med. 1997 Jun 15;126(12):946-54
references.pmid: 9182471
references.citation: Clin Cancer Res. 2007 Feb 1;13(3):920-8
references.pmid: 17289886
references.citation: J Biomed Opt. 2008 May-Jun;13(3):034008
references.pmid: 18601553
references.citation: J Biomed Opt. 2007 Sep-Oct;12(5):051503
references.pmid: 17994867
references.citation: Nat Med. 1999 Jan;5(1):83-9
references.pmid: 9883844
references.citation: Annu Rev Med. 2013;64:31-44
references.pmid: 23092385
references.citation: Blood. 2009 Apr 30;113(18):4341-51
references.pmid: 19139079
references.citation: Opt Lett. 2006 Dec 15;31(24):3623-5
references.pmid: 17130924
references.citation: Opt Express. 2007 Jul 9;15(14):8681-94
references.pmid: 19547203
references.citation: Chem Biol. 2009 Nov 25;16(11):1169-79
references.pmid: 19942140
references.citation: Cancer Treat Rev. 2009 Aug;35(5):463-74
references.pmid: 19410375