Moss gametophyte
Longitudinal cross section of an antheridial head on gametophyte stage of a moss (I think, I am still learning.) This is where the sperm cells are produced. Specimen from the edge of the Dowagiac River (5/2/15).
The following protocol is largely based on Chamberlain, "Methods in Plant Histology, 5th ed., 1905. Fixed 24 hours in aqueous chromic acid-acetic acid (3.7 g potassium dichromate/l + 10 ml glacial acetic acid/l). (The substation of potassium dichromate for chromic acid based on equivalent chromate conc. is reported to work as long as the pH is less than 3.4). Washed 24 hours in flowing water. Dehydrated in 10, 35, 50, 75, 85, 95, 99 % IPA in water, 6 hours each min. Infiltrated in xylene saturated with Paraplast for 2 days, followed by 2 changes of melted Praplast for 2 hours each. Embedded in Paraplast. Sectioned on a Spencer 820 microtome at 11 micron. Cleared in Xylene 2X, 10 min each. Rehydrated 99 (10 min), 95, 85, 70 % IPA, 2 min. each. Stained Gill's Hematoxylin 10 sec. Washed 3 min running water. Blued 0.05 % lithium carbonate 30 s. Water rinse 1 min. Stained 1 % aq. Eosin-Y 1 min. Dehydrated 99 % IPA 2 min. Cleared 2X xyene 5 min each. Mounted with DEPEX.
Photographed on a Spencer 42 petrographic polarizing microscope using an original magnification of , using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.
Moss gametophyte
Longitudinal cross section of an antheridial head on gametophyte stage of a moss (I think, I am still learning.) This is where the sperm cells are produced. Specimen from the edge of the Dowagiac River (5/2/15).
The following protocol is largely based on Chamberlain, "Methods in Plant Histology, 5th ed., 1905. Fixed 24 hours in aqueous chromic acid-acetic acid (3.7 g potassium dichromate/l + 10 ml glacial acetic acid/l). (The substation of potassium dichromate for chromic acid based on equivalent chromate conc. is reported to work as long as the pH is less than 3.4). Washed 24 hours in flowing water. Dehydrated in 10, 35, 50, 75, 85, 95, 99 % IPA in water, 6 hours each min. Infiltrated in xylene saturated with Paraplast for 2 days, followed by 2 changes of melted Praplast for 2 hours each. Embedded in Paraplast. Sectioned on a Spencer 820 microtome at 11 micron. Cleared in Xylene 2X, 10 min each. Rehydrated 99 (10 min), 95, 85, 70 % IPA, 2 min. each. Stained Gill's Hematoxylin 10 sec. Washed 3 min running water. Blued 0.05 % lithium carbonate 30 s. Water rinse 1 min. Stained 1 % aq. Eosin-Y 1 min. Dehydrated 99 % IPA 2 min. Cleared 2X xyene 5 min each. Mounted with DEPEX.
Photographed on a Spencer 42 petrographic polarizing microscope using an original magnification of , using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.