Vascular bundles in a Petiole Gall
I collected this petiole gall from a yet to be identified bush near the Dowagiac River. This is the same gall as illustrated in www.flickr.com/photos/14643312@N02/20281620511. This section is cut normal to where the gall attached to the petiole and it shows the primary vascular bundles entering the gall.
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.
Vascular bundles in a Petiole Gall
I collected this petiole gall from a yet to be identified bush near the Dowagiac River. This is the same gall as illustrated in www.flickr.com/photos/14643312@N02/20281620511. This section is cut normal to where the gall attached to the petiole and it shows the primary vascular bundles entering the gall.
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.