several antibodies that can be used on such tissue. DAPI labelling of nuclei cannot
be used on Epon embedded tissue; when this is required, embedding in LRWhite
resin can be used instead (Fig. 6.9). Additionally, secondary antibodies conjugated
to both a fluorophore and colloidal gold can be applied to further confirm the
correspondence between immunofluorescence and ultrastructure (Fig. 6.9).
Table 6.2 Antibodies that can be used on conventionally prepared tissue for EM (including
osmium postfixation and Epon embedding)
Antigen
Host
Antibody source
RRID
GABA
Guinea pig
Millipore AB175
RRID:AB_91011
GABA
Rabbit
Millipore AB131
RRID:AB_2278931
Glutamate
Rabbit
Millipore AB5018
RRID:AB_91640
a Tubulin
Rabbit
Abcam ab18251
RRID:AB_2210057
Acetylated a tubulin
Mouse
Sigma T6793
RRID:AB_477585
Glutamine synthetase
Mouse
BD Biosciences 610517
RRID:AB_397879
Fig. 6.9 Conjugate AT on conventionally prepared EM tissue embedded in LRWhite. GABA
(green) immunostaining and DAPI labeling of ultrathin sections from mouse cortex fixed with 2%
formaldehyde and 2.5% glutaraldehyde, postfixed with 1% OsO 4 and 1.5% K ferricyanide, and
embedded in LRWhite. GABA antibody (Millipore AB131) was used at 1:100 dilution followed
by a goat anti-rabbit Alexa 488–10 nm colloidal gold conjugate at 1:50
6 Conjugate Immunofluorescence—SEM Array Tomography …
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