Coprinellus disseminatus
This is a dark field image of an oblique section through the cap (top left) into a gill of Coprinellus disseminatus mushroom. The spore producing basidia are stained red while the mycelium is yellow green. Note that the basidia extend across the underside of the cap in a continuous fashion to the neighboring gill. The specimen was from a cluster under a tree near the Dowagiac River.
The following protocol for fixation is largely based on Chamberlain, "Methods in Plant Histology, 5th ed., 1905. Staining was per Johansen. Fixed 48 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:paraplast 3:1, 1:1, 1:3 1 hour each. Infiltrated in xylene saturated with Paraplast for 10 days, followed by 2 changes of melted Paraplast for 2 hours each. Embedded in Paraplast. Sectioned on a Spencer 820 microtome at 5 micron. Cleared in Xylene 2X, 10 min each. Rehydrated 99 (10 min), 90 (10 min.), 85, 70 % IPA, 2 min. each. Stained Gill's Hematoxylin 20 sec. Washed 3 min running water. Blued 0.05 % lithium carbonate 10 s. Water rinse 1 min. Stained 1 % aq. Erythrosin-B 2 min. Dehydrated 99 % IPA 2 min 2X. Cleared 2X xylene 5 min each. Mounted with DEPEX.
Photographed on a Spencer 43 petrographic microscope with a spencer oil immersion dark field condenser using an original magnification of 430X using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.
Coprinellus disseminatus
This is a dark field image of an oblique section through the cap (top left) into a gill of Coprinellus disseminatus mushroom. The spore producing basidia are stained red while the mycelium is yellow green. Note that the basidia extend across the underside of the cap in a continuous fashion to the neighboring gill. The specimen was from a cluster under a tree near the Dowagiac River.
The following protocol for fixation is largely based on Chamberlain, "Methods in Plant Histology, 5th ed., 1905. Staining was per Johansen. Fixed 48 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:paraplast 3:1, 1:1, 1:3 1 hour each. Infiltrated in xylene saturated with Paraplast for 10 days, followed by 2 changes of melted Paraplast for 2 hours each. Embedded in Paraplast. Sectioned on a Spencer 820 microtome at 5 micron. Cleared in Xylene 2X, 10 min each. Rehydrated 99 (10 min), 90 (10 min.), 85, 70 % IPA, 2 min. each. Stained Gill's Hematoxylin 20 sec. Washed 3 min running water. Blued 0.05 % lithium carbonate 10 s. Water rinse 1 min. Stained 1 % aq. Erythrosin-B 2 min. Dehydrated 99 % IPA 2 min 2X. Cleared 2X xylene 5 min each. Mounted with DEPEX.
Photographed on a Spencer 43 petrographic microscope with a spencer oil immersion dark field condenser using an original magnification of 430X using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.