PMID- 32925887
OWN - NLM
STAT- MEDLINE
DCOM- 20201127
LR  - 20201127
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 162
DP  - 2020 Aug 26
TI  - Successful In vivo Calcium Imaging with a Head-Mount Miniaturized Microscope in 
      the Amygdala of Freely Behaving Mouse.
LID - 10.3791/61659 [doi]
AB  - In vivo real-time monitoring of neuronal activities in freely moving animals is 
      one of key approaches to link neuronal activity to behavior. For this purpose, an 
      in vivo imaging technique that detects calcium transients in neurons using 
      genetically encoded calcium indicators (GECIs), a miniaturized fluorescence 
      microscope, and a gradient refractive index (GRIN) lens has been developed and 
      successfully applied to many brain structures(1) (,) (2) (,) (3) (,) (4) (,) (5) 
      (,) (6). This imaging technique is particularly powerful because it enables 
      chronic simultaneous imaging of genetically defined cell populations for a 
      long-term period up to several weeks. Although useful, this imaging technique has 
      not been easily applied to brain structures that locate deep within the brain 
      such as amygdala, an essential brain structure for emotional processing and 
      associative fear memory(7). There are several factors that make it difficult to 
      apply the imaging technique to the amygdala. For instance, motion artifacts 
      usually occur more frequently during the imaging conducted in the deeper brain 
      regions because a head-mount microscope implanted deep in the brain is relatively 
      unstable. Another problem is that the lateral ventricle is positioned close to 
      the implanted GRIN lens and its movement during respiration may cause highly 
      irregular motion artifacts that cannot be easily corrected, which makes it 
      difficult to form a stable imaging view. Furthermore, because cells in the 
      amygdala are usually quiet at a resting or anesthetized state, it is hard to find 
      and focus the target cells expressing GECI in the amygdala during baseplating 
      procedure for later imaging. This protocol provides a helpful guideline for how 
      to efficiently target cells expressing GECI in the amygdala with head-mount 
      miniaturized microscope for successful in vivo calcium imaging in such a deeper 
      brain region. It is noted that this protocol is based on a particular system 
      (e.g., Inscopix) but not restricted to it.
FAU - Lee, Han-Sol
AU  - Lee HS
AD  - Department of Biological Sciences, Korea Advanced Institute of Science and 
      Technology (KAIST); KAIST Institute for BioCentury (KIB), Korea Advanced 
      Institute of Science and Technology (KAIST); science226@kaist.ac.kr.
FAU - Han, Jin-Hee
AU  - Han JH
AD  - Department of Biological Sciences, Korea Advanced Institute of Science and 
      Technology (KAIST); KAIST Institute for BioCentury (KIB), Korea Advanced 
      Institute of Science and Technology (KAIST).
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20200826
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Calcium-Binding Proteins)
RN  - 0 (GCaMP2)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Acoustic Stimulation
MH  - Amygdala/*diagnostic imaging
MH  - Animals
MH  - Artifacts
MH  - Behavior, Animal/*physiology
MH  - Calcium/*metabolism
MH  - Calcium-Binding Proteins/metabolism
MH  - Green Fluorescent Proteins/metabolism
MH  - Head
MH  - Lenses
MH  - Mice, Inbred C57BL
MH  - Microscopy/*instrumentation
MH  - Miniaturization/*instrumentation
MH  - Movement
MH  - Neuroimaging
MH  - Neurons/metabolism
MH  - Refractometry
MH  - Reproducibility of Results
MH  - Stereotaxic Techniques
EDAT- 2020/09/15 06:00
MHDA- 2020/11/28 06:00
CRDT- 2020/09/14 15:45
PHST- 2020/09/14 15:45 [entrez]
PHST- 2020/09/15 06:00 [pubmed]
PHST- 2020/11/28 06:00 [medline]
AID - 10.3791/61659 [doi]
PST - epublish
SO  - J Vis Exp. 2020 Aug 26;(162). doi: 10.3791/61659.