Modified Hiraoka TEM grid staining apparatus and technique using 3D printed materials and gadolinium triacetate tetrahydrate, a nonradioactive uranyl acetate substitute

Abstract The Hiraoka transmission electron microscopy (TEM) grid staining apparatus, which allowed multiple grids to be batch stained together on a plate, is no longer commercially available. Because the need for such a device still exists, we developed a modified Hiraoka TEM grid staining apparatus by combining a Leica AC20 grid plate holder with a modified 3D printed grid loading holder and trays lined with ParafilmR. This apparatus was then used to test a post-section contrasting method that uses gadolinium triacetate tetrahydrate solution, a nonradioactive uranyl acetate substitute, which produces similar staining results to uranyl acetate when used with lead stain in mouse ocular tissue for TEM.

[1]  P. Seifert,et al.  Astrocytes in the Optic Nerve Head of Glaucomatous Mice Display a Characteristic Reactive Phenotype , 2017, Investigative ophthalmology & visual science.

[2]  Steve D. M. Brown,et al.  Mouse Models of NMNAT 1-Leber Congenital Amaurosis ( LCA 9 ) Recapitulate Key Features of 77 78 79 80 81 82 the Human Disease , 2016 .

[3]  Antony Bacic,et al.  Preparation of plant cells for transmission electron microscopy to optimize immunogold labeling of carbohydrate and protein epitopes , 2012, Nature Protocols.

[4]  E. Katayama,et al.  New versatile staining reagents for biological transmission electron microscopy that substitute for uranyl acetate. , 2011, Journal of electron microscopy.

[5]  S. Kanematsu,et al.  Enhanced effects of nonisotopic hafnium chloride in methanol as a substitute for uranyl acetate in TEM contrast of ultrastructure of fungal and plant cells , 2010, Microscopy research and technique.

[6]  A. Adachi,et al.  Oolong tea extract as a substitute for uranyl acetate in staining of ultrathin sections , 2008, Journal of microscopy.

[7]  H. Nakane,et al.  Platinum blue as an alternative to uranyl acetate for staining in transmission electron microscopy. , 2007, Archives of histology and cytology.

[8]  J. Syväoja,et al.  Device for handling electron microscopy grids. , 2006, BioTechniques.

[9]  B. Trump,et al.  A method for staining epoxy sections for light microscopy. , 1961, Journal of ultrastructure research.

[10]  J I Hiraoka,et al.  A holder for mass treatment of grids, adapted especially to electron staining and autoradiography. , 1972, Stain technology.