Developing sporocytes
This is an image of the early stage of sporocyte development in a moss sporophyte. This is indicated by the angular shape and the observation that they have pulled away from the cell walls. (K.S. Renzaglia, K.D. McFarland, D.K. Smith, American J. Botany, [84] (10) 1337-1350 (1997).) Longitudinal cross section of a developing sporophyte capsule. A photo of the moss from which the sporophytes were obtained is at www.flickr.com/photos/14643312@N02/17164418555/. A complete sporophyte is in www.flickr.com/photos/14643312@N02/17173980525/. Stained with Flemming's Safranin, Gentian Violet, Orange Stain, substituting Crystal Violet for Gentian Violet. This specimen was sectioned at 7 micron.
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 Paraplast 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 % IPA,10 min. each. Stained 1% Safranin in 50% IPA 24 hr. Differentiated 50 % IPA, 10 min. Stained 1 % aq Crystal Violet 10 min. Dip 4X in water. Stain 1 % Orange B 1 min. Dip 4X 95 % IPA, agitate 5 sec 99 % IPA. Rinse off IPA with Eugenol, wick off. Pour Eugenol over slide. Cleared 2X xylene 2.5 min each. Mounted with DEPEX.
Bright field photomicrograph from a Nikon MS inverted microscope at 400X original magnification using a Leica MIKAS 1/3X adapter to a Sony NEX 5N.
Developing sporocytes
This is an image of the early stage of sporocyte development in a moss sporophyte. This is indicated by the angular shape and the observation that they have pulled away from the cell walls. (K.S. Renzaglia, K.D. McFarland, D.K. Smith, American J. Botany, [84] (10) 1337-1350 (1997).) Longitudinal cross section of a developing sporophyte capsule. A photo of the moss from which the sporophytes were obtained is at www.flickr.com/photos/14643312@N02/17164418555/. A complete sporophyte is in www.flickr.com/photos/14643312@N02/17173980525/. Stained with Flemming's Safranin, Gentian Violet, Orange Stain, substituting Crystal Violet for Gentian Violet. This specimen was sectioned at 7 micron.
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 Paraplast 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 % IPA,10 min. each. Stained 1% Safranin in 50% IPA 24 hr. Differentiated 50 % IPA, 10 min. Stained 1 % aq Crystal Violet 10 min. Dip 4X in water. Stain 1 % Orange B 1 min. Dip 4X 95 % IPA, agitate 5 sec 99 % IPA. Rinse off IPA with Eugenol, wick off. Pour Eugenol over slide. Cleared 2X xylene 2.5 min each. Mounted with DEPEX.
Bright field photomicrograph from a Nikon MS inverted microscope at 400X original magnification using a Leica MIKAS 1/3X adapter to a Sony NEX 5N.