Asarum canadense bright field
This is a bright field transverse section through the stem at the base of a leaf (dorsal side down) of Asarum canadense, or wild ginger showing the vascular structures. This is a bright field view of the polarized light image in: www.flickr.com/photos/14643312@N02/17547325725. Time in Safranin-O was increased from 2 to 24 hr. This made differentiation of the regressive dye much easier to control. Safranin-O stains nucle and lignin red and Fast Green stains cytoplasm green. Specimen collected along the Dowagiac River, 25 May 2015.
The protocol was as follows. Specimens fixed in FAA (formaldehyde, acetic acid, ethanol) 48 hr. Dehydrated in IPA at 35, 50, 70, 85, 91, 95, 99 %. Infiltrated with paraplast saturated xylene 48 hr followed by 3 Paraplast baths prior to embedding in Paraplast. Sectioned 7 um thick on a Spencer 820 microtome. Cleared in xylene, 5 min, 2X. Rehydrated in 99, 95, 80, 70 %IPA. Stained in Johansen's Safranin-O, Fast Green (24 hours in Safranin-O, 15 sec. in fast green). Cleared 2 X in xylene 10 min each and mounted with DPEX.
Photographed in bright field on a Spencer 42 petrographic microscope at original magnification using a Sony NEX-5N and a Leica MIKAS 1/3X adapter.
Asarum canadense bright field
This is a bright field transverse section through the stem at the base of a leaf (dorsal side down) of Asarum canadense, or wild ginger showing the vascular structures. This is a bright field view of the polarized light image in: www.flickr.com/photos/14643312@N02/17547325725. Time in Safranin-O was increased from 2 to 24 hr. This made differentiation of the regressive dye much easier to control. Safranin-O stains nucle and lignin red and Fast Green stains cytoplasm green. Specimen collected along the Dowagiac River, 25 May 2015.
The protocol was as follows. Specimens fixed in FAA (formaldehyde, acetic acid, ethanol) 48 hr. Dehydrated in IPA at 35, 50, 70, 85, 91, 95, 99 %. Infiltrated with paraplast saturated xylene 48 hr followed by 3 Paraplast baths prior to embedding in Paraplast. Sectioned 7 um thick on a Spencer 820 microtome. Cleared in xylene, 5 min, 2X. Rehydrated in 99, 95, 80, 70 %IPA. Stained in Johansen's Safranin-O, Fast Green (24 hours in Safranin-O, 15 sec. in fast green). Cleared 2 X in xylene 10 min each and mounted with DPEX.
Photographed in bright field on a Spencer 42 petrographic microscope at original magnification using a Sony NEX-5N and a Leica MIKAS 1/3X adapter.