荧光成像和荧光检测.pdf

荧光成像

孙晓莲
xiaolian-******@
Optical Properties of NPs
Beer-Lambert Law

Elastic scattering -εcL
A = ln( I / I0) = ε CL I = I0 e
λ2 = λ1
Where A is the absorbance.

ε is the molar absorption/ the extinction coefficient
Incident
C is the concentration of the chemical in the cuvette
Photon λ1
Absorption ( in molar, M).
E = hc/λ
L is the path length (cm) of the cuvette.

I is the intensity (energy per unit area) getting
through the solution.
Emitted photon λ2 > λ1:

Luminescence
I0 is the light intensity applied to the solution.
Fluorescence
Phosphorescence The Beer-Lambert law states that absorbance is linear upon
Raman scattering concentration (at low concentrations).
Basic Concepts
• Luminescence: the emission of light by a
materials as consequence of its absorbing
energy. Various types of excitation source
may be used.
• Photoluminescence:uses photos or light,
often UV for excitation.
• Electroluminescence: uses an electrical
energy input

• For a short time lapse, ~< 10-8 s, between
excitation and emission, the process is
known as fluorescence. Fluorescence
effectively ceases as soon as the excitation
sou