All cells were cultured under identical conditions and the experiments were performed with a minimum of three biological replicates (cell preparations from different hemangioma specimens) and three technical replicates.
In fact, immunohistochemistry is semiquantitative and highly dependent on a range of variables, such as choice of antibody, antibody concentration, fixation techniques, variability in the interpretation and stratification criteria, and inconsistency in specimen handling and technical procedures.
However, these methods require refrigeration, a cold chain for specimen transport, and technical expertise; therefore, they are often performed only in reference/diagnostic labs far removed from patients, potentially limiting their clinical utility.
Amongst others, we conducted an in-vitro study ourselves, using 10 human knee specimens and the same technical parameters [ 39].
Although the MMIHC approach contains a sampling process within a whole section of tissue, it is obvious that an assay that could accurately quantify several cancer-related proteins simultaneously on single tumor section or small tumor specimens does offer clear advantages and technical advances over standard immunohistochemical method [21].
The authors thank Dr. Preetha Ramalingam for help with Pathology Review of the Specimens, and Vasanta Putluri for technical assistance.
There are several reasons that specimen coordinates might be at variance with the Jepson eFlora including: the need to revise range statements in the Jepson eFlora, incorrect coordinates assigned to specimens, misidentifications or unincorporated taxonomic revisions, and technical mapping issues.
For both countries, ticks positive for Rickettsia spp. and SFG rickettsiae DNA were male and female specimens (Technical Appendix Table).
Thanks to the University of Washington Dermatology Research Center for providing facilities for electron microscopy specimen preparation, especially to Robert Underwood for training and technical advice.
