Fig. 4: CaBLAM shows SNR comparable to GCaMP under epifluorescent illumination during in vivo mammalian imaging. | Nature Methods

Fig. 4: CaBLAM shows SNR comparable to GCaMP under epifluorescent illumination during in vivo mammalian imaging.

From: CaBLAM: a high-contrast bioluminescent Ca2+ indicator derived from an engineered Oplophorus gracilirostris luciferase

Fig. 4

a, The mouse imaging setup with location of the cranial window and head-fixation on a running wheel. b, The CaBLAM and GCaMP6s imaging configurations: CaBLAM was imaged in darkness in a light-tight enclosure after administration of luciferin and GCaMP6s was imaged under epifluorescent illumination. c, Mean projection images of an NDNF-Cre CaBLAM and GCaMP6s field of view. d, CaBLAM activity traces from 15 neurons in 1 field of view. Vertical gray bars indicate onset of tactile stimuli, and the (bottom) trace is mouse running speed. e, Percentages of stimulus responsive cells across animals and sensor groups. Individual mice are represented by dots and group medians by horizontal bars. f, Example of 20 tactile stimulation trials for 4 cells from 1 field of view. Gray traces are individual trials, and black traces are the mean. g, Mean responses of all positively responsive cells for the NDNF CaBLAM (left) and NDNF GCaMP6s (right) groups sorted in ascending order by latency of the signal at half-maximum. h, Mean sensory response across mice in the two groups from positively responsive cells. Stimulation onset is at t = 0, indicated by the vertical dashed line. Opaque bars represent the jackknifed 95% CI of the means. Separate left and right y axes for the CaBLAM and GCaMP6s signals, respectively. i, Single-trial SNR measurements across all positively responsive cells for the two sensor groups. Dots are individual trials and horizontal lines indicate the median. j, Mean running-onset waveforms across animals for the two sensor groups. Running onset is at t = 0, indicated by the vertical dashed line. Inset, the average running speed across mice time-locked to onset of running bouts. k, Left: single epifluorescence image of a full cranial window expressing CaBLAM pan-neuronally in one animal. Right: the CaBLAM BL average projection image (~4 min recording, n = 5,119 images) in the same mouse after retro-orbital injection of CFz. l, Mean CaBLAM tactile response (n = 3 mice) after retro-orbital injections of CFz (sampled ROI shown by the white circle in k). Stimulation onset is at t = 0, indicated by the vertical dashed line. For the 2-Hz sampling rate (magenta), opaque shading represents the jackknifed 95% CI of the mean. The darker trace shows an average tactile response from one animal imaged at 10 Hz.

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