EBs from different time points (day 6.5 to 10.5) were washed with PBS and embedded in OCT freezing compound (Miles Scientific, Cambridge, ML) after fixation in 4% paraformaldehyde for 1 hour at 4°C and cryoprotection in 20% (wt/vol) sucrose overnight.
For immunohistochemistry, excised corneal samples were embedded in frozen section compounds (Surgipath; Leica Microsystems, Nussloch, Germany), and stored at −80 °C until sectioning.
Last year, the Smiths lost nearly all their wheat to a late freeze compounded by too much rain.
Another anti-freeze compound, recently discovered in an Alaskan beetle that can tolerate -60C, might prove more useful.
Wedge liver biopsies were obtained at the time of portoenterostomy and snap-frozen in liquid nitrogen, embedded frozen in OCT compound, or formalin fixed/paraffin embedded.
Alternatively, samples were frozen in OCT compound (Tissue Tek), and frozen sections were labeled with anti-K5 (AF138, Covance), anti-K8 (Troma-I, Developmental Studies Hybridoma Bank), anti-CD11b, anti-Gr-1 (ebioscience) or Ulex Europaeus Agglutinin I (UAE, Vector Laboratories).
Spleen sections were frozen in OCT compound (Tissue-Tek), and frozen sectioned (10 μm thick) using a cryostat (Leica).
The brains were removed, cryoprotected in 30% sucrose, and frozen in O.C.T. compound (Sakura Finetek, Torrance, CA, USA).
The target was a saline solution of EGFR or EGR, the guest compound, frozen by thermal contact with liquid nitrogen.
To qualitatively measure the distribution of the GNPs as well as to provide a measure of MCL growth characteristics, MCL inserts were frozen in OCT compound for sectioning.
