Theoretical Background: Fluorescence Imaging
Theoretical Background
Fluorescence
Fluorescence is the ability of many natural and synthetic structures, known as fluorophores, to emit light when excited with light at specific wavelength. Fluorescence phenomena occur when an incident light photon interacts with an electron of a fluorophore. The incident light photon may transfer its energy to the electron of the fluorophore and hence the electron moves to a higher energy state. When the electron returns to its ground state, it loses energy which is known as fluorescence.
Fluorescence imaging
Fluorescence Imaging is being used in biological sciences with aim to visualize cells and tissues in vitro and in vivo. It has many benefits, including high sensitivity because it can be often seen
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The stability of the spectrum is fastest when the ICG is dissolved in distilled water. Furthermore, the near-infrared absorption peak in tissues and cells moves to longer wavelength because of the cell proteins binding.
Optical Properties
ICG rapidly bounds to plasma protein, when it is injected into the human body and generates fluorescence in near infrared at about 800 nm and longer wavelengths when is excited by a light source between 750-800nm. The fluorescence spectrum of ICG depends on the its concentration, the temperature and on the chemical environment. Likewise, ICG fluorescence spectrum smoothly varying with the light from the excitation source and the filters, which are used.
Penetration
Hemoglobin and water are the main optical absorbers in the human tissue. Hemoglobin absorbs the visible light below the wavelength of 650nm and the water absorbs the infrared light above the wavelength of 900nm. ICG works in the optical window which is the infrared wavelength between 650nm and 900nm and it is relatively 'transparent' because the absorption of hemoglobin and water is low. For this reason, ICG when is injected into human skin, is able to observe vascular properties several millimeters or ever further (about 10mm) from the surface of the skin
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