Panaeolus foenisecii gill structure
This is an oblique section through the gills of a Panaeolus foenisecii mushroom cut near the cap. The whole mushroom is shown at www.flickr.com/photos/14643312@N02/18306291616. The basidia which produce the spores at their tips are the dark blue stained structures at the right. The spores are the ovoid orange structures. Moving to the left below the basidia the mycelium which make up the structure of the gills is composed of hyphae, i.e., the strands. This is the same material as the vegetative structure in the substrate on which the mushroom fruiting body grows. The isolated spores in the mycelium are artifacts of specimen preparation. This is not the best staining job; 20 sec was too long in Hematoxylin, however, it does show the general teardrop shape of the basidia. Note how they constrict before joining to the mycelium.
The following protocol for fixation is largely based on Chamberlain, "Methods in Plant Histology, 5th ed., 1905. Staining was per Johansen. 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. Cleared IPA:Xylene, 1:3, 1:1, 3:1, 2 hr. 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 20 sec. Washed 3 min running water. Blued 0.05 % lithium carbonate 3 s. Water rinse 1 min. Stained 1 % aq. Erythrosin-B 2 min. Dehydrated 99 % IPA 2 min 2X. Cleared 2X xyene 5 min each. Mounted with DEPEX.
Photographed on a Spencer 42 petrographic polarizing microscope using an original magnification of 430X using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.
Panaeolus foenisecii gill structure
This is an oblique section through the gills of a Panaeolus foenisecii mushroom cut near the cap. The whole mushroom is shown at www.flickr.com/photos/14643312@N02/18306291616. The basidia which produce the spores at their tips are the dark blue stained structures at the right. The spores are the ovoid orange structures. Moving to the left below the basidia the mycelium which make up the structure of the gills is composed of hyphae, i.e., the strands. This is the same material as the vegetative structure in the substrate on which the mushroom fruiting body grows. The isolated spores in the mycelium are artifacts of specimen preparation. This is not the best staining job; 20 sec was too long in Hematoxylin, however, it does show the general teardrop shape of the basidia. Note how they constrict before joining to the mycelium.
The following protocol for fixation is largely based on Chamberlain, "Methods in Plant Histology, 5th ed., 1905. Staining was per Johansen. 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. Cleared IPA:Xylene, 1:3, 1:1, 3:1, 2 hr. 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 20 sec. Washed 3 min running water. Blued 0.05 % lithium carbonate 3 s. Water rinse 1 min. Stained 1 % aq. Erythrosin-B 2 min. Dehydrated 99 % IPA 2 min 2X. Cleared 2X xyene 5 min each. Mounted with DEPEX.
Photographed on a Spencer 42 petrographic polarizing microscope using an original magnification of 430X using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.