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The patient labels were fixed as positive class labels.
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But this is not possible in the standard multi-label formulation because it is transductive the labels are fixed during the training phase, and predictions on new labels are not possible.
Cells for immunogold labeling were fixed with ice cold 4% paraformaldehyde in PBS for 1 h at room temperature followed by blocking and permeablization with 5% goat serum/0.1% saponin for 1 h.
For immunostaining and Tdt-mediated dUTP nick-end labeling (TUNEL) labeling, OVs were fixed for 2 h in 4% (w/v) paraformaldehyde (Merck Chemicals, Mollet del Valles, Spain) at 4 °C.
The same procedure was used for live cell labeling; after sequential incubation and labeling, cells were fixed with 3% PFA, and nuclei were counterstained with DAPI.
The labeled cells were fixed with 4% paraformaldehyde and stained with DAPI.
After being washed with PBS, the labeled cells were fixed with Cytofix/Cytoperm solution and stained with DAPI in PBS.
labeling, cells were fixed, permeabilized and blocked as described above.
For BCθ labeling, cells were fixed with 4% PFA and stained with BCθ and Alexa594-conjugated streptavidin as described [15].
For mMHC-B labeling, cells were fixed in 4% formaldehyde in PBS for 20 minutes and blocked with PBST.
In brief, labeled cells were fixed with 4% paraformaldehyde and images obtained with a Leica DM IRE2 microscope [66], [67].
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