C57 b6 mice

Male BALB/c mice and C57/B6 mice (6–8-week of age) were purchased from the Shanghai Experimental Animal Center (Shanghai, China). Animals were housed in pathogen-free animal cages under a 12-h light/dark cycle with free access to food and water. The experimental procedures were approved by the Animal Ethic Committee at Tongji University and Shenzhen University.

The current study was approved by the Animal Ethics Committee of Shanghai Medical College, Fudan University, Shanghai, China (ID:20160225-013 and 20200306-004). Twenty-six adult female nude mice and 21 C57B/6 mice were purchased from Shanghai Laboratory Animal Center (SLAC) (Shanghai, China). The mice were housed at 5 or 6 animals per cage under controlled standard conditions (21–23 °C, 34–40% humidity, and a 12 h light/12 h dark cycle). All the mice had free access to tap water, and they were fed the same standard laboratory chow (SLAC, Shanghai, China). Twenty-six nude mice were randomly divided into two groups, and eight were used for the in vitro CUBIC experiment while 18 were used for the in vivo NIR-II experiment (Fig. 1). For the CUBIC experiment, mice were assigned to the control group and the EA group (n = 4). In the in vivo NIR-II experiment, 18 mice were evenly and randomly distributed into the control, MA, and EA groups (n = 6). The twenty-one C57B/6 mice were randomly divided into the control (n = 7), ovariectomy (OVX, n = 7), and ovariectomy with EA (OVX + EA, n = 7) groups.Fig. 1

Experimental methodology flow. EA, electroacupuncture; MA, manual electroacupuncture; OVX, ovariectomy; CUBIC, clear, unobstructed brain/body imaging cocktails and computational analysis; H&E staining, hematoxylin and eosin staining.