Paraphyses
This is part of the Paraphyses of a moss. These single cell wide hairs surround the female gametophyte in the moss. They contain chloroplasts as shown here.
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, min 6 hours each. Cleared in IPA:Xylene at 3:1, 2:1, 1:3,1 hour each and pure xylene for 10 min. Infiltrated in xylene saturated with Paraplast for 5 days, followed by 3 changes of melted Praplast for 2 hours each. Embedded in Paraplast. Sectioned on a Spencer 820 microtome at 7 micron. Cleared in Xylene 2X, 10 min each. Rehydrated 99, 95, 70 % IPA, followed by water for 5-10 min each. Stained in Gills Haematoxylin, 5 sec, water washed 5 min. Stained 1% Eosin, 1 min. Dehydrated 2X 99% IPA, 2 MIN. Cleared 2X xyene 2.5 min each. Mounted with DEPEX.
Paraphyses
This is part of the Paraphyses of a moss. These single cell wide hairs surround the female gametophyte in the moss. They contain chloroplasts as shown here.
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, min 6 hours each. Cleared in IPA:Xylene at 3:1, 2:1, 1:3,1 hour each and pure xylene for 10 min. Infiltrated in xylene saturated with Paraplast for 5 days, followed by 3 changes of melted Praplast for 2 hours each. Embedded in Paraplast. Sectioned on a Spencer 820 microtome at 7 micron. Cleared in Xylene 2X, 10 min each. Rehydrated 99, 95, 70 % IPA, followed by water for 5-10 min each. Stained in Gills Haematoxylin, 5 sec, water washed 5 min. Stained 1% Eosin, 1 min. Dehydrated 2X 99% IPA, 2 MIN. Cleared 2X xyene 2.5 min each. Mounted with DEPEX.