Systems and methods for recording simultaneously visible light image and infrared light image from fluorophores

An imaging system for imaging a sample comprising an infrared or near-infrared fluorophore, comprising:

• an image sensor;
• a laser to emit an excitation light for the infrared or near-infrared fluorophore, wherein the excitation light excites the infrared or near-infrared fluorophore in the sample to emit an emission light;
• a notch filter in the emission light path from the sample to the image sensor, wherein the notch filter blocks a portion of the excitation light reflected from the sample towards the image sensor;
• a white light source to emit a light comprising visible light, wherein the visible light is conducted to the sample, wherein the sample reflects the visible light, and wherein the reflected visible light is conducted to the image sensor;
• and
• an image processing unit to process signals from the image sensor to generate a white light frame (WLF) when the sample receives only visible light, a stray light frame (SLF) when the sample receives neither visible light nor the excitation light, and two or more near infrared frames (NIFs) when the sample receives only the excitation light to reduce a ghosting effect.

A method for imaging a sample comprising an infrared or near-infrared fluorophore, comprising:

• operating a laser to emit an excitation light for the infrared or near-infrared fluorophore;
• operating a notch filter in the emission light path from the sample to the single image sensor to block a portion of the excitation light reflected from the sample towards the single image sensor;
• operating a white light source to emit a light comprising visible light;
• conducting the visible light to the sample, wherein the sample reflects the visible light;
• conducting the reflected visible light to the single image sensor;
• and
• operating an image processing unit to process signals from the single image sensor to generate a white light frame (WLF) when the sample receives only visible light, a stray light frame (SLF) when the sample receives neither visible light nor the excitation light, and two or more near infrared frames (NIFs) when the sample receives only the excitation light to reduce a ghosting effect.

An imaging system for imaging a sample comprising an infrared or near-infrared fluorophore, comprising:

• an image sensor;
• a laser to emit an excitation light for the infrared or near-infrared fluorophore, wherein the excitation light excites the infrared or near-infrared fluorophore in the sample to emit an emission light;
• a laser clean-up filter in the light path from the laser to the sample;
• a white light source to emit a light comprising visible light, wherein the visible light is conducted to the sample, wherein the sample reflects the visible light, wherein the reflected visible light is conducted to the image sensor;
• and
• a notch beam splitter in the light path from the laser to the sample and in the light path from the white light source to the sample, whereby the excitation light is reflected by the notch beam splitter to the sample and the visible light is transmitted by the notch beam splitter to the sample.
• an image processing unit to process signals from the image sensor to generate a white light frame (WLF) when the sample receives only visible light, a stray light frame (SLF) when the sample receives neither visible light nor the excitation light, and two or more near infrared frames (NIFs) when the sample receives only the excitation light to reduce a ghosting effect.

A method for imaging a sample comprising an infrared or near-infrared fluorophore, comprising:

• operating a laser to emit an excitation light for the infrared or near-infrared fluorophore;
• operating a white light source to emit a light comprising visible light;
• conducting the visible light to the sample, wherein the sample reflects the visible light;
• operating a notch beam splitter in the light path from the laser to the sample and in the light path from the white light source to the sample;
• conducting the reflected visible light to the single image sensor;
• and
• operating an image processing unit to process signals from the single image sensor to generate a white light frame (WLF) when the sample receives only visible light, a stray light frame (SLF) when the sample receives neither visible light nor the excitation light, and two or more near infrared frames (NIFs) when the sample receives only the excitation light to reduce a ghosting effect.