Confocal microscopy of thick sections from acrylamide gel embedded embryos

Microscopy Research and Technique
P G GermrothR P Thompson

Abstract

The preparation of optically clear, thick sections of fragile embryonic tissues greatly aids the power of confocal scanning laser microscopy in imaging three-dimensional structures. We report here conditions for embedding, sectioning, and staining embryos in polyacrylamide gels for a variety of confocal imaging techniques. Infiltration of tissues in standard mixtures of 10-15% acrylamide monomer yields, upon polymerization, blocks that cut easily by vibratome between 50 and 1,000 microns. These conditions worked well for tissues previously stained or for staining gel sections with low molecular weight water-soluble fluorochromes (MW < 5 kD [e.g., propidium iodide, phalloidin]). For immunostaining of tissue after embedding and sectioning, the acrylamide concentration was reduced to 2-3% acrylamide to allow access of immunoglobulins to antigenic sites; such gels were supplemented with 1% agarose to facilitate sectioning and handling. Either method yielded abundant, optically clear, and easily handled sections for mounting and examination in water-miscible media.

References

Sep 1, 1991·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·D D BronsonW E Stumpf
Nov 1, 1987·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·T H Rosenquist, J R McCoy
Jan 1, 1989·The Anatomical Record·H SumidaR P Thompson
Apr 1, 1984·Developmental Biology·W Halfter, S Deiss
Aug 1, 1993·Journal of Toxicology and Environmental Health·B S Jortner, M Ehrich

❮ Previous
Next ❯

Citations

Mar 29, 2000·The Anatomical Record·D SedmeraR H Anderson
Sep 20, 2000·The Anatomical Record·N HuE B Clark
May 17, 2012·Journal of Microbiological Methods·Fiona J BairdJane E Hill
Aug 3, 2005·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·David SedmeraRobert P Thompson
Aug 3, 2005·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Christine E MillerDavid Sedmera
Nov 27, 2002·Journal of Anatomy·J L SchibV Sans-Coma
Feb 24, 2001·Progress in Histochemistry and Cytochemistry·G Mayer, M Bendayan
Jan 29, 2005·The Anatomical Record. Part A, Discoveries in Molecular, Cellular, and Evolutionary Biology·Kimimasa TobitaBradley B Keller
May 7, 2009·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Brooke J DamonRobert P Thompson
Oct 20, 2007·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Tim C McQuinnDavid Sedmera
Jul 30, 2011·The Journal of Comparative Neurology·Scott J SchachteleSteven H Green
Mar 23, 2000·Mechanisms of Development·E Frolova, D Beebe
Aug 5, 2008·Developmental Biology·Donald BellFernando Giraldez
Nov 9, 2017·Frontiers in Computational Neuroscience·Young-Gyu YoonEdward S Boyden
Jul 18, 2017·Nature Biotechnology·Yongxin ZhaoEdward S Boyden
Jun 26, 2018·The FEBS Journal·Shahar AlonEdward S Boyden
Sep 16, 2004·The Anatomical Record. Part A, Discoveries in Molecular, Cellular, and Evolutionary Biology·David SedmeraRobert P Thompson
Aug 3, 2018·Current Protocols in Cell Biology·Shoh M AsanoEdward S Boyden
Jul 14, 2005·Journal of Experimental Zoology. Part A, Comparative Experimental Biology·Josè M IcardoBruno Tota
May 9, 2006·American Journal of Physiology. Heart and Circulatory Physiology·David SedmeraRobert G Gourdie
Aug 20, 2019·Annual Review of Cell and Developmental Biology·Paul W Tillberg, Fei Chen
Sep 29, 2020·Frontiers in Cell and Developmental Biology·Zsuzsa AkosAndras Czirok
Jun 21, 2017·BMC Biology·Ruixuan GaoEdward S Boyden
Jul 12, 2003·American Journal of Physiology. Heart and Circulatory Physiology·Christine E MillerDavid Sedmera
May 13, 2017·The Journal of Cell Biology·Cataldo SchietromaChristine Petit
Mar 22, 2001·Archives of Pathology & Laboratory Medicine·A H BoagM J Miller
Dec 23, 2004·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Yumei ChenAnn F Ramsdell
Oct 25, 2017·Current Opinion in Biomedical Engineering·Yu Shrike ZhangAli Khademhosseini

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cell Adhesion Molecules in the Brain

Cell adhesion molecules found on cell surface help cells bind with other cells or the extracellular matrix to maintain structure and function. Here is the latest research on their role in the brain.