Emcu ultra microtome

The LGs were first fixed in a solution of 3% glutaraldehyde (#01909-10, Polysciences, Warrington, PA, USA) and then washed in a 1 M sodium phosphate buffer (PBS) with a pH of 7.3. They were subsequently post-fixed in an osmium tetroxide (OsO4; #0972A-20, Polysciences, Warrington, PA, USA) solution for one hour. After fixation, the samples underwent dehydration through a series of graded alcohols, acetone, and Polybed 812 plastic resin (#08792-1, Polysciences, Warrington, PA, USA). Finally, they were embedded in plastic block molds using 100% Polybed 812. One-micron sections were obtained using a Leica EMCU ultra microtome and placed on glass slides. The sections were then stained with toluidine blue (#01234-25, Polysciences, Warrington, PA, USA).

Testis samples were fixed in 3% glutaraldehyde. Tissue samples were washed in 1 M sodium phosphate buffer (pH 7.3), post fixed in 1% osmium tetroxide for 1 h, and dehydrated through graded alcohol washes. Tissue samples were infiltrated (hardened) with acetone and Polybed 812 plastic resin and embedded in plastic block molds with 100% Polybed 812. One-micron sections (thick sections) were cut on an ultra-Leica EMCU ultra microtome and placed on glass slides and stained with Toluidine Blue. Ultra-thin Sects. (80 nm) were cut from sample blocks using a Leica EMUC Ultra microtome and mounted on 100 mesh copper grids. Grids were stained with 2% uranyl acetate and Reynold’s lead stain. Grids were analyzed on a JEOL JEM 1250 electron microscope and images were captured on an AMTV600 digital camera.