Cortical Calcium Imaging of GCaMP6 Fluorescence

WFI was performed on day 1or 2 of the training step 3 (beginner) and on day 21 or 22 (expert) using a CMOS camera (FLASH4; C-13 440-20CU; Hamamatsu) system, adapted from a previous study (Kim et al., 2016 (link)). The cortical surface was imaged through a 2×/0.08 NA objective (PlanApo N; Olympus). For measurement of GCaMP6 emission, a 470 nm LED (Optogenetics-LED-Blue; Prizmatrix) was connected to the dichroic cube holder using a 1000-μm-diameter 0.50-NA fiber (M59L01; Thorlabs). The light was filtered with a 470 nm bandpass filter (FB470-10; Thorlabs) that was fiber-coupled into the dichroic mirror (FF495-Di03-25 × 36; Semrock). Fluorescence emitted from the cortical surface was passed through a 530 nm bandpass fluorescence emission filter (FF01-520/35-25; Semrock). Images were acquired using HCImage Live (Hamamatsu) at 20 Hz (50 ms/frame) and 192 × 192 pixels. Imaging was started by using the transistor-transistor logic (TTL) signal generated from the Arduino to synchronize the imaging and behavior data.

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Jeong M., Lee H., Kim Y., Wang E.H., Paik S.B., Lim B.K, & Kim D. (2021). Interhemispheric Cortico-Cortical Pathway for Sequential Bimanual Movements in Mice. eNeuro, 8(4), ENEURO.0200-21.2021.

Publication 2021

PlanApo N FB470-10 FF495-Di03-25 × 36 FF01-520/35-25 HCImage Live

Cmos Cortical Fiber Fluorescence Frame Light Optogenetics

Corresponding Organization :

Other organizations : University of California, San Diego, Korea Advanced Institute of Science and Technology

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Variable analysis

independent variables

Day of training step (1 or 2, and 21 or 22)

dependent variables

GCaMP6 emission

control variables

CMOS camera (FLASH4; C-13 440-20CU; Hamamatsu)
2×/0.08 NA objective (PlanApo N; Olympus)
470 nm LED (Optogenetics-LED-Blue; Prizmatrix)
Fiber-coupled 470 nm bandpass filter (FB470-10; Thorlabs)
Dichroic mirror (FF495-Di03-25 × 36; Semrock)
530 nm bandpass emission filter (FF01-520/35-25; Semrock)
Image acquisition at 20 Hz (50 ms/frame) and 192 × 192 pixels using HCImage Live (Hamamatsu)
Synchronization of imaging and behavior data using TTL signal from Arduino

controls

Positive control not mentioned
Negative control not mentioned

Annotations

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