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New en bloc staining technique improve tissue contrast and electrical conductivity (1050.2)
Author(s) -
NegronMorales Celimar,
TorresVazquez Irma,
SerranoVelez Jose,
Powell Richard,
RosaMolinar Eduardo
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1050.2
Subject(s) - epoxy , materials science , electrical conductor , shrinkage , staining , biomedical engineering , conductivity , composite material , chemistry , pathology , medicine
Here we describe results of our work to develop a conductive resin and en bloc staining technique that together overcome the challenges of “charging” and “electron beam damage” posed by three‐dimensional (3D) electron‐based imaging methods such as serial‐section electron tomography. A conductive epoxy resin optimized for infiltration, immunocytochemistry, and uniform serial‐sectioning did not infiltrate evenly throughout the tissue and did not section well; however, it provided tissue section stability and eliminated tissue section charging, beam damage, and shrinkage. This led to our development of a new en bloc staining technique to use with the epoxy resin. Prior to embedding tissue in a non‐conductive resin, we made the tissue‐electron‐conductive and non‐charging. Together, the resin and the staining technique increase tissue contrast and electrical conductivity and overcome challenges posed by charging, radiation, and ion beam‐and electron‐induced surface contamination/artifacts. New tools such as the ones described here are needed to overcome the challenges of new technologies and take advantage of the increased resolution they provide. Supported by NS‐080687; GM‐108470; MH‐086994; NSF‐1062963, and NSF‐0964114.