Systems and methods for recording simultaneously visible light image and infrared light image from fluorophores
A fluorophore, visible-light technology for use in analysis, applications, diagnostic recording/measurement, etc. using fluorescence emission
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Embodiment 1
[0148] Charge-coupled device (CCD) or complementary metal-oxide-semiconductor (CMOS) sensors used in cameras have broad-spectrum sensitivities in the 400nm to 1000nm range ( figure 2 ). All red, green and blue sensors showed sensitivities in the wavelength range of 800-1000nm. Commercially available cameras have on top of the sensor such as image 3 The color filter array (CFA) or color filter mosaic (CFM) used to capture the color information of an image as shown in . In addition to this filter array there is an additional NIR short pass filter that cuts off light at wavelengths of 700-1000 nm.
Embodiment 2
[0150] We use the sensitivity of red, green and blue pixels in the near-infrared region (NIR) to detect infrared fluorescence. A visible light source illuminates the sample of interest. In addition, a laser is used as the excitation light for the infrared fluorophore in the tissue, and the emitted light of the infrared fluorophore is detected by a CCD camera. At the same time, the excitation light is filtered before reaching the CCD camera so as not to interfere with the detection of the emitted light. Image frames are captured when the laser is on (on frame). Another image frame is captured while the laser is off (off frame). Both visible and infrared fluorescence is detected on the frame, rather than only visible light on the frame. Thus, the difference in intensity between on-frame and off-frame provides information about the infrared fluorescence signal. ( Figure 4 ).
[0151] 1. To stimulate:
[0152] Excitation is achieved using a very narrow wavelength laser at ...
Embodiment 3
[0158] Fluorescence emitted by NIR fluorophores can be detected on all RGB channels by removing the NIR short-pass filter in front of the sensor ( figure 2 ). But in order to differentiate between visible and NIR light, we have to make sure that no visible light is present on the sensor when the NIR image frame is captured. In order to capture NIR light, no visible light should be present. In some cases, when no visible or NIR light is present, we capture a frame, record the light and then subtract that frame from the NIR captured frame. Clinical prototypes are shown in Figures 6A-6C middle.
[0159] 1. Filter combination:
[0160] We use very specific filter combinations to achieve the highest signal-to-noise ratio (SNR). Instead of using broadband excitation as in most current NIR systems, we use an extremely narrow band excitation at 785nm (optimal for ICG, may vary depending on the fluorophore), which is further narrowed using a laser-cleaned filter ( Figure 7 ), ...
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