Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Jan 22;11:612678. doi: 10.3389/fphys.2020.612678

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2021 Kılıç, Desjardins, Tang, Thunemann, Sunil, Erdener, Postnov, Boas and Devor.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

(A) Schematics of the borosilicate glass window implant. (B) Schematic illustration of the window implant over the whisker representation within the primary somatosensory cortex (SI) and the headpost fixed to the skull overlaying the other (contralateral) hemisphere. (C) Images of the brain vasculature through the glass window implant obtained by 2-photon imaging of fluorescein isothiocyanate (FITC)-labeled dextran injected intravenously. The images illustrate preserved integrity of the vasculature between days 1 (left) and 28 (right) following surgical implantation. (D) Two-photon image stack obtained with Alexa 680 labeled dextran injected intravenously illustrating the capability of deep imaging. The resolution for presented images was ~1 μm/pixel. (E) Top: Image of the surface vasculature calculated as a maximum intensity projection (MIP) of an image stack 0–300 μm in depth using a 4 × objective. Individual images were acquired every 10 μm (top, left). A zoomed-in view of the region within the red square acquired with a 20× objective (top, middle, left). A plane 250 μm below the surface corresponding to the region outlined in blue (top, middle, right). The yellow circle indicates a small diving arteriole. An example temporal diameter change profile acquired from the arteriole outlined by the yellow circle imaged 250 μm below the surface (top, right). The vessel diameter was captured by repeated line-scans across the vessel. These line-scans form a space-time image when stacked sequentially, from left to right. White arrowheads indicate the onset of stimulus trials (air puffs to the whisker pad); four trials are shown. Graphs: Single-vessel dilation time-courses extracted from data. Time-courses for individual trials are overlaid for sensory stimuli (left, n = 160 trials) and OG stimuli (right, n = 19 trials); the thick lines show the average. The stimulus onset is indicated by the black arrowhead and the blue vertical line for the sensory and OG panels, respectively. (F) Concurrent IOSI and LSCI in the same subject. A CCD reflectance image of the surface vasculature (left). The corresponding LS contrast image (middle, left). Ratio images of HbO (extracted from the OIS data, see Methods) and LS contrast showing the region of activation following OG stimulation (middle, right, and right). The location of optical fiber is indicated on all images (black dotted line). The same arteriole is outlined by yellow circles. The black parallelogram indicates the region of interest (ROI) used for extraction of time-courses. The resolution for presented images was 5.5 μm/pixel. Graphs: Time-courses of HbO and HbR (shown in red and blue, respectively), and LS contrast (shown in black) in response to sensory and OG stimulation. These time-courses were extracted from the polygonal ROI shown in top images. (G) Corrected GE EPI image (left, top) and a corresponding structural image (TurboRARE, left, bottom). The resolution for EPI and TurboRARE is 200 μm/pixel 100 × 50 and ~75 μm/pixel, slice thickness = 1 mm. Red arrows point to the peripheral edges of the implant, i.e., the glass/bone boundary. The red line indicates the bottom of the glass implant, i.e., the glass/brain boundary. The BOLD signal in response to sensory stimuli in a fully awake mouse (top, middle). Spatiotemporal evolution of the BOLD signal change from a single slice cutting through the center of the evoked response, presented as trial-averaged ratio maps, in response to a 20-s train of 100-ms light pulses delivered at 1 Hz (“blocked” OG stimulus) in a single Emx1-Cre/Ai32 subject. EPI images were thresholded to reflect the sensitivity of the surface RF coil (for display purposes only). The ratio images are overlaid on the structural (TurboRARE) image of the same slice. BOLD response time-courses extracted from the active ROI (top, right). Fifty seven stimulus trials are superimposed. The average is overlaid in thick black. The BOLD signal in response to OG stimuli in a fully awake mouse (bottom, middle). Spatiotemporal evolution of the BOLD signal change from a single slice cutting through the center of the evoked response, presented as trial-averaged ratio maps, in response to a 20-s train of 100-ms light pulses delivered at 1 Hz (“blocked” OG stimulus) in a single Emx1-Cre/Ai32 subject. EPI images were thresholded to reflect the sensitivity of the surface RF coil (for display purposes only). The ratio images are overlaid on the structural (TurboRARE) image of the same slice. BOLD response time-courses extracted from the active ROI (bottom, right). Twenty eight stimulus trials are superimposed. The average is overlaid in thick black. Modified from Desjardins et al. (2019).