Techniques / Tools

Multi-Photon Time Lapse In Vivo Imaging

The Cruz-Martín lab has a 2P-microscope
adapted for in vivo imaging

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In vivo imaging of layer 5 pyramidal neurons in the somatosensory cortex (labeled using the Thy1-GFP mouse

3D-printed miniature microscopes for in vivo imaging in freely moving mice

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Vasoactive Intestinal Polypeptide (VIP) - expressing interneurons in the cingulate expressing GCaMP6f, a calcium indicator

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Imaging the activity of VIP-expressing interneurons in the cingulate while mice perform a social behavioral task

We can use our imaging methods to monitor the activity of
hundreds of cells

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We are currently using DeepLabCut to track mice while they perform a
behavioral task (Mathis et al. 2018; Johnson and Kretsge et al. 2020)

We can use this approach to track multiple mice at the same time

We are studying the role of immune genes in cortical synaptic development. During early postnatal development, some synapses are eliminated, in part, via microglia-dependent synaptic phagocytosis. We use a method called Expansion microscopy (ExM, from Ed Boyden's group) to visualize this biological process.

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We can visualize synapses using FingRs (Fibronectin intrabodies generated with mRNA display), that bind endogenous neuronal proteins PSD-95 (Gross et al. 2013). Below are labeled synapses in a dendrite (Comer et al. 2020).

We can identify different neuronal subtypes using multiplex in situ hybridization (M-FISH, RNAScope). Here we stain for parvalbumin and somatostatin in the prefrontal cortex of a mouse

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We can dissect the connectivity patterns of moleculary identified brain cell types using simultaneous whole-cell patch-clmap recordings.

IMARIS reconstruction based on STED microscopy. Using this nano-resolution approach we identified PSD95 (green) and Rab11a (magenta) puncta that are within 0.2 um from the border of the spines.