An improved method for three-dimensional reconstruction of protein expression patterns in intact mouse and chicken embryos and organs

被引：64

作者：

Ahnfelt-Ronne, Jonas ^{[1
]}

Jorgensen, Mette C. ^{[1
]}

Hald, Jacob ^{[1
]}

Madsen, Ole D. ^{[1
]}

Serup, Palle ^{[1
]}

Hecksher-Sorensen, Jacob ^{[1
]}

机构：

[1] Hagedorn Res Inst, Dept Dev Biol, DK-2820 Gentofte, Denmark

来源：

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY | 2007年 / 55卷 / 09期

关键词：

wholemount; immunofluorescence; confocal; pancreas; three-dimensional; imaging;

D O I：

10.1369/jhc.7A7226.2007

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We have developed a whole mount immunofluorescence protocol for the simultaneous detection of up to three proteins in mouse and chicken embryos. Combined with Murray's clearing reagent (BABB) and microscope objectives with long working ranges and high numerical apertures mounted on a confocal microscope, cellular resolution can be obtained in depths offering the possibility of examining expression patterns in entire organs or embryos. Three-dimensional projections of the optical confocal sections can be computed with computer software allowing rotation around any axis. The protocol is robust and we find that most antibodies working on tissue sections also work with this protocol. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.

引用

页码：925 / 930

页数：6

共 17 条

[1] Tomographic molecular imaging and 3D quantification within adult mouse organs [J].

Alanentalo, Tomas ;

Asayesh, Amir ;

Morrison, Harris ;

Loren, Christina E. ;

Holmberg, Dan ;

Sharpe, James ;

Ahlgren, Ulf .

NATURE METHODS, 2007, 4 (01) :31-33

[2] Requirement for the homeobox gene Hb9 in the consolidation of motor neuron identity [J].

Arber, S ;

Han, B ;

Mendelsohn, M ;

Smith, M ;

Jessell, TM ;

Sockanathan, S .

NEURON, 1999, 23 (04) :659-674

[3] Notch ligands with contrasting functions: Jagged1 and Delta1 in the mouse inner ear [J].

Brooker, R ;

Hozumi, K ;

Lewis, J .

DEVELOPMENT, 2006, 133 (07) :1277-1286

[4] FISHing for chick genes:: Triple-label whole-mount fluorescence in situ hybridization detects simultaneous and overlapping gene expression in avian embryos [J].

Denkers, N ;

García-Villalba, P ;

Rodesch, CK ;

Nielson, KR ;

Mauch, TJ .

DEVELOPMENTAL DYNAMICS, 2004, 229 (03) :651-657

[5]

DENT JA, 1989, DEVELOPMENT, V105, P61

[6] Fgf10 regulates hepatopancreatic ductal system patterning and differentiation [J].

Dong, P. Duc Si ;

Munson, Chantilly A. ;

Norton, William ;

Crosnier, Cecile ;

Pan, Xiufang ;

Gong, Zhiyuan ;

Neumann, Carl J. ;

Stainier, Didier Y. R. .

NATURE GENETICS, 2007, 39 (03) :397-402

[7] neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas [J].

Gradwohl, G ;

Dierich, A ;

LeMeur, M ;

Guillemot, F .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (04) :1607-1611

[8]

Gu GQ, 2002, DEVELOPMENT, V129, P2447

[9] Direct lineage tracing reveals the ontogeny of pancreatic cell fates during mouse embryogenesis [J].

Gu, GQ ;

Brown, JR ;

Melton, DA .

MECHANISMS OF DEVELOPMENT, 2003, 120 (01) :35-43

[10] A SERIES OF NORMAL STAGES IN THE DEVELOPMENT OF THE CHICK EMBRYO [J].

HAMBURGER, V ;

HAMILTON, HL .

JOURNAL OF MORPHOLOGY, 1951, 88 (01) :49-&

← 1 2 →