Petiole Gall
I collected a petiole gall from a yet to be identified bush along the dowagiac River. Its internal structure consisted of several chambers such as shown here, each with what would appear to be a single un identified insect larva. The Gall was C-shaped and open to the center and each cavity was close to/connected to this open centre. In this case, the cutting plane does not reveal this, however, the opening was to the lower left. The cells approaching the cavity became progressively smaller and more dense, having a structure similar to the epithelial cells on the exterior of the gall. On a upward diagonal from lower left to upper right, the cells making up the gall take on the character of leaf mesophyll. A leaf from this plant has been embedded but not yet sectioned for comparison.
The protocol was as follows. Specimens fixed in FAA (formaldehyde, acetic acid, ethanol) 24 hr. Dehydrated in 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 11 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 Nikon MS inverted microscope using an original magnification of 40X, using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.
Petiole Gall
I collected a petiole gall from a yet to be identified bush along the dowagiac River. Its internal structure consisted of several chambers such as shown here, each with what would appear to be a single un identified insect larva. The Gall was C-shaped and open to the center and each cavity was close to/connected to this open centre. In this case, the cutting plane does not reveal this, however, the opening was to the lower left. The cells approaching the cavity became progressively smaller and more dense, having a structure similar to the epithelial cells on the exterior of the gall. On a upward diagonal from lower left to upper right, the cells making up the gall take on the character of leaf mesophyll. A leaf from this plant has been embedded but not yet sectioned for comparison.
The protocol was as follows. Specimens fixed in FAA (formaldehyde, acetic acid, ethanol) 24 hr. Dehydrated in 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 11 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 Nikon MS inverted microscope using an original magnification of 40X, using a Sony NEX 5N with a Leica MIKAS 1/3X adapter.