Calcium Imaging of AM Dyes Following Prolonged Incubation in Acute Neuronal Tissue
Fig 1
Calcium imaging in acute brain slices A) Flow diagram illustrating the slicing and calcium AM dye loading protocol in brain slices. B) Fluorescence images of a neocortical slice shows ubiquitous staining of cortical neurons and glia with Fura-2-AM (left image; 380nm excitation; 20x objective; scale bar 100 μm). Application of 30 mM KCl, caused a noticeable increase in 340/380nm ratio relative to F0 (dF/F), depicting three classes of cells according to their staining pattern. Left–Loaded cells; Middle–Spontaneously active cells (imaged as ratiometric changes before the application of KCl) and Right–Evoked cells (following 30mM KCl). Red and blue correspond to high and low Ca2+ concentrations, respectively C) Bar graph depicting the percentage of spontaneous and evoked cells, out of the total loaded cells within the field of view indicating a slight yet insignificant decrease in the both spontaneous and evoked cells following >24 hours in the Braincubator (p>0.9; two tailed student t-test).