At present, one of the most interesting light sources for microscopy is the light-emitting diode (LED), which underwent significant engineering efforts in the last two decades. High-power LEDs are commercially selleck Bicalutamide available, and could be thought of as compact and inexpensive sources for single molecule detection and sensing applications, or even for single photon generation. Their availability across the spectral range from 280 to 1,300 nm, their stable output, their electrical insensitivity, and long lifetime make them attractive alternatives to laser diodes for some applications.In this paper we discuss the use of a commercially available LED as an excitation source for single molecule studies. Unlike presented before [7,8], we extend the experiments to the green part of the visible spectrum.
The Inhibitors,Modulators,Libraries minimum irradiance to excite and detect single molecules is estimated by comparing the expected illumination levels to the nominal sensitivity of a camera and single photon detectors. A rigorous proof that a single molecule has been observed is only possible by detecting the characteristic anti-bunched photon statistics . Prospects Inhibitors,Modulators,Libraries for the generation and detection of single photons based on LED illumination are discussed. Experimentally, we compare LEDs and laser excitation schemes side by side. It is shown that single molecules can be imaged with LED illumination Inhibitors,Modulators,Libraries and the influencing factors are presented. This extends the work of Kuo and coworkers, which mention present experimental findings on single molecule detection in the blue part of the spectrum .2.
?Material and MethodsThe sample for all further experiments and estimations consists of a doped thin crystalline film of p-terphenyl (Aldrich) which is spin coated on a microscopy coverslip . As a dopant, the well characterized fluorescent Inhibitors,Modulators,Libraries molecule terrylene was chosen [11,12]. The concentration of terrylene molecules (PAH Research) is in the order of ��10?10 to allow spatial separation. In an area of 10 �� 10 ��m2 1�C20 molecules were observed. The coverslips were cleaned by organic solvents in an ultrasonic bath and afterwards transferred to a solution of 1 part sulfuric acid and 3 parts hydrogen peroxide Brefeldin_A (pira?a solution) to clean organic residues. The coverslips continue to be submerged in the solution for storage until use and rinsed under water prior to spin coating sample preparation.All our single molecule studies were performed on a custom-made inverted microscope which can be configured for wide field or confocal imaging, and selleckchem can use LED or laser illumination in either configuration (Figure 1). As a well characterized light source a frequency doubled Nd:YAG laser (532 nm) was coupled into the excitation path via a single mode optical fiber.