Journal: bioRxiv
Article Title: Axially decoupled photo-stimulation and two photon readout ( ADePT ) for mapping functional connectivity of neural circuits
doi: 10.1101/2025.02.24.639992
Figure Lengend Snippet: (a) (Top) Cartoon schematics of functional connectivity mapping of a neural circuit (e.g. olfactory bulb); photo-stimulation (2-D light mask from a digital micro-mirror device illuminated with a solid-state CW laser) and two photon imaging are confined to different optical z-planes that can be flexibly and independently adjusted by translating the diffuser and respectively the primary objective; (Bottom) Alternating (strobing) between photo-stimulation and imaging periods. Each red bar represents a single frame of multiphoton imaging. Photo-stimulation and imaging periods are interleaved. ( b) Microscope schematics. DM, dichroic mirrors. DMD, digital micro-mirror device. I, iris diaphragm. L1-L12, lenses. O, primary objective. PMT, photomultiplier tube. PS, periscope. S, shutter. SM, scan mirrors. ( c) (Top) Illustration of using a movable diffuser to decouple the patterned photo-stimulation and multiphoton imaging planes. The diffuser is imaged into the sample, in a 4f lens configuration; translating the diffuser along the optical path causes the corresponding projection plane to shift axially. OFP, objective focal plane. PSP, photo-stimulation plane. (d) DMD chip-to-CCD camera-to-2p microscope registration. (i) two photon micrograph of 10 µm fluorescence microbeads; arrows mark two microbeads, part of the larger DMD-modulated projection target pattern (8 microbeads), which were taken as fiduciary points; (ii) widefield fluorescence image (full field illumination) of a larger field of view including the target microbeads; (iii) ROIs selected from the 2p image were used to generate DMD-chip light masks; these were further projected at the primary objective focal plane and imaged using the primary CCD camera (CCD 1); (iv) overlay of DMD-generated photo-stimulation masks and widefield fluorescence image of 10 µm microbeads from (ii); note that fluorescence is restricted to the microbeads targeted by the DMD photo-stimulation masks with minimal spillover to adjacent (off-target) microbeads (see fiduciary marks).
Article Snippet: Patterned photo-stimulation is achieved using a digital micro-mirror device (VX4100 DMD, 0.7” XGA VIS 1024 x 768 pixels, 13.68 μm micro-mirror pitch, Vialux) illuminated by a solid-state CW laser (Sapphire, 488 nm, 200 mW, Coherent).
Techniques: Functional Assay, Olfactory, Imaging, Microscopy, Fluorescence, Generated
Coherent Corp is a verified supplier 
Coherent Corp manufactures this product 
