anti mouse igg  (Biotium)

Journal: Nature Methods

Article Title: Dense 4D nanoscale reconstruction of living brain tissue

doi: 10.1038/s41592-023-01936-6

Figure Lengend Snippet: GCaMP recording, chemogenetically activated single bouton. Full series of the GCaMP6f recording shown in Fig. 5a . Ca 2+ transients of a mossy fiber bouton attached to a hilar mossy cell in Prox1-cre::Ai95 mouse organotypic hippocampal slice cultures are visible. LIONESS (greyscale) and dTomato (confocal, orange, coexpressed with the DREADD hM3Dq) images are identical replicates placing the overlaid time-varying Ca 2+ signals (confocal, green) after stimulation with the DREADD ligand CNO into structural context. Acquisition frame rate of the GCaMP signal was 2 Hz. LIONESS images are maximum intensity projections spanning 150 nm. Edge lengths, 14.8 µm × 14.4 µm.

Article Snippet: Chemically targeted activation with simultaneous calcium imaging of neurons was conducted using AAVs containing a Cre-dependent DREADD , construct (AAV-DIO-CAG-hM3Dq-2A-dTomato; Addgene #202036) added to organotypic hippocampal slice cultures of Prox1-cre::Ai95 (GCaMP6f) mice at 4–6 d in vitro.

Techniques:

Journal: Nature Methods

Article Title: Dense 4D nanoscale reconstruction of living brain tissue

doi: 10.1038/s41592-023-01936-6

Figure Lengend Snippet: a , Single plane of an isotropically super-resolved LIONESS volume in the hilus of the DG in an organotypic hippocampal slice culture, where a subset of mossy fiber boutons expressed both the excitatory DREADD hM3Dq (together with cytosolic dTomato, confocal, purple, virally delivered) and the calcium indicator GCaMP6f (confocal, green, Prox1-cre::Ai95 mouse). LIONESS and dTomato images are identical replicates, placing the overlaid time-varying Ca 2+ signals after stimulation with the DREADD ligand CNO into structural context, showing three example points from a time series. Note that confocal signals extend beyond the structures defined by LIONESS. In particular, they partially originate from structures above and below the plane displayed here. Inset shows GCaMP signal (averaged pixel value normalized to first frame) as a function of time. LIONESS image is a maximum intensity projection spanning 150 nm. Scale bar, 2 µm. b , 3D reconstructions of a hM3Dq-expressing mossy fiber bouton (purple) and its postsynaptic partner, a hilar mossy cell (gold) with complex spines at two time points (top, day 0 (0 h); bottom, day 1 (19.5 h)). V 0h and V 19.5h are bouton volumes at the respective time points. Green (i) and blue (ii) frames indicate the viewing angles from opposite directions for the magnified views on the right. The structures designated by the lettering in a and b refer to the same bouton and complex spine. The scale cube refers to the center of the rendering. Scale bars in the magnified views correspond to 1 µm in the center of the respective bouton renderings.

Article Snippet: Chemically targeted activation with simultaneous calcium imaging of neurons was conducted using AAVs containing a Cre-dependent DREADD , construct (AAV-DIO-CAG-hM3Dq-2A-dTomato; Addgene #202036) added to organotypic hippocampal slice cultures of Prox1-cre::Ai95 (GCaMP6f) mice at 4–6 d in vitro.

Techniques: Expressing