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Journal: PLoS ONE
Article Title: In Situ Optical Mapping of Voltage and Calcium in the Heart
doi: 10.1371/journal.pone.0042562
Figure Lengend Snippet: ( A ) di-4-ANBDQPQ fluorescence in a Langendorff-perfused rat heart (sinus rhythm), excited with blue (blue LED, 470±10 nm filter), green (green LED, 540±12.5 nm filter) and red (red LED, 640±10 nm filter) wavelengths. These fluorescence signals (taken from the 4×4-pixel white-square region on the left-ventricle) were collected through a custom-made multi-band emission filter (F3 in B , C ). The green trace ([Ca 2+ ] i ) shows negligible emission changes when di-4-ANBDQPQ is excited at the excitation-isosbestic point. Scale bar = 5 mm. ( B ) Schematic outline of the imaging system, highlighting key components. Since only one camera is used, the system requires no challenging optical alignment. Excitation sources Ex1: red LED with a 640±10 nm filter (F1), Ex2: green LED with a 540±12.5 nm filter (F2). ( C ) Transmission spectrum of the custom multi-band emission filter that passes both V m (Em1) and [Ca 2+ ] i (Em2) emitted fluorescence signals. F1 and F2 excitation filter spectra are shown as dashed curves. ( D ) Basic principle behind the single-camera multi-LED approach: During any frame exposure (occurring between the vertical dashed lines), the parameter being measured by the camera sensor is determined by the excitation source (either Ex1 or Ex2) that is switched on during that period. A sufficiently fast camera (compared to Em1 and Em2 signal dynamics), and interpolation between measured data points, provides simultaneous measures of multiple parameters, here V m and [Ca 2+ ] i . For these images, blebbistatin was used to eliminate the contribution of motion to the signals.
Article Snippet: Fluorescence emission from dye-loaded hearts was passed through custom-made
Techniques: Fluorescence, Imaging, Transmission Assay