In Vivo Three-Dimensional Two-Photon Microscopy to Study Conducted Vascular Responses by Local ATP Ejection Using a Glass Micro-Pipette
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In Vivo Three-Dimensional Two-Photon Microscopy to Study Conducted Vascular Responses by Local ATP Ejection Using a Glass Micro-Pipette. / Cai, Changsi; Zambach, Stefan A.; Fordsmann, Jonas C.; Lonstrup, Micael; Thomsen, Kirsten J.; Jensen, Aske G. K.; Lauritzen, Martin.
In: Journal of Visualized Experiments, Vol. 148, e59286, 2019.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - In Vivo Three-Dimensional Two-Photon Microscopy to Study Conducted Vascular Responses by Local ATP Ejection Using a Glass Micro-Pipette
AU - Cai, Changsi
AU - Zambach, Stefan A.
AU - Fordsmann, Jonas C.
AU - Lonstrup, Micael
AU - Thomsen, Kirsten J.
AU - Jensen, Aske G. K.
AU - Lauritzen, Martin
PY - 2019
Y1 - 2019
N2 - Maintenance of normal brain function requires a sufficient and efficient supply of oxygen and nutrition by a complex network of vessels. However, the regulation of cerebral blood flow (CBF) is incompletely understood, especially at the capillary level. Two-photon microscopy is a powerful tool widely used to study CBF and its regulation. Currently, this field is limited by the lack of in vivo two-photon microscopy studies examining (1) CBF responses in three-dimensions, (2) conducted vascular responses, and (3) localized interventions within the vascular network. Here, we describe a 3D in vivo method using two-photon microscopy to study conducted vascular responses elicited by local ejection of ATP with a glass micro-pipette. Our method uses fast and repetitive hyperstack two-photon imaging providing precise diameter measurements by maximal intensity projection of the obtained images. Furthermore, we show that this method can also be used to study 3D astrocytic calcium responses. We also discuss the advantages and limitations of glass micro-pipette insertion and two-photon hyperstack imaging.
AB - Maintenance of normal brain function requires a sufficient and efficient supply of oxygen and nutrition by a complex network of vessels. However, the regulation of cerebral blood flow (CBF) is incompletely understood, especially at the capillary level. Two-photon microscopy is a powerful tool widely used to study CBF and its regulation. Currently, this field is limited by the lack of in vivo two-photon microscopy studies examining (1) CBF responses in three-dimensions, (2) conducted vascular responses, and (3) localized interventions within the vascular network. Here, we describe a 3D in vivo method using two-photon microscopy to study conducted vascular responses elicited by local ejection of ATP with a glass micro-pipette. Our method uses fast and repetitive hyperstack two-photon imaging providing precise diameter measurements by maximal intensity projection of the obtained images. Furthermore, we show that this method can also be used to study 3D astrocytic calcium responses. We also discuss the advantages and limitations of glass micro-pipette insertion and two-photon hyperstack imaging.
KW - Neuroscience
KW - Issue 148
KW - in vivo
KW - two-photon microscopy
KW - cerebral blood flow (CBF)
KW - astrocytes
KW - three dimensions (3D)
KW - neurovascular coupling
KW - calcium imaging
KW - viral vector injection
U2 - 10.3791/59286
DO - 10.3791/59286
M3 - Journal article
C2 - 31233031
VL - 148
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
SN - 1940-087X
M1 - e59286
ER -
ID: 225380392