Panaeolus foenisecii mushroom
This is a transverse section through the fruiting body of a Panaeolus foenisecii mushroom showing the gills and the spores. The whole mushroom is shown at www.flickr.com/photos/14643312@N02/18306291616.
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 100X using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.
Panaeolus foenisecii mushroom
This is a transverse section through the fruiting body of a Panaeolus foenisecii mushroom showing the gills and the spores. The whole mushroom is shown at www.flickr.com/photos/14643312@N02/18306291616.
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 100X using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.