This molecule can already exist within the specimen (endogenous or auto-fluorescence), be applied externally and attached to the constituent (chemically or through antibody binding), or transfected (fluorescent proteins) into the cell.In order for the molecule to emit light (fluoresce) it must first absorb light (a photon) with the appropriate amount of energy to promote the molecule from the ground state to the excited state, as seen in Figure 3A below. The amount of fluorescence is proportional to the intensity (I) of the incident laser, and so confocal LSM is often referred to as a linear imaging technique. Third harmonic generation (THG) is observed at boundary interfaces where there is a refractive index change. could you please explain easily?Hello, thank you for contacting Thorlabs. This means that if we take our 60X objective from earlier, the lateral resolution is 249 nm (Equation 2) and the pixel size in the displayed image should be 108 nm. The spot size is the distance between the zeros of the Airy disk (diameter across the middle of the first ring around the center of the bullseye) and is termed one Airy Unit (AU). A pinhole of 1 AU is a good balance between signal strength, resolution, and confocality.In multiphoton LSM, a short pulsed free-space laser supplies the collimated illumination beam that passes through the scanning system and is focused by the objective.

The included ThorImageAll hardware components are directly controlled by the ThorImageLS software, including automated Z-step control for optical sectioning (via a piezo or stepper motor) and automatic calculation of Airy disk units based on the combination of the objective magnification and pinhole size. This means that the resolution of the optical system can be improved by reducing the size of the pinhole. The standard sensitivity multi-alkali PMTs provide a low-noise image with high dynamic range that is appropriate for most life-science and industrial applications. The amount of fluorescence is proportional to the intensity (I) of the incident laser, and so confocal LSM is often referred to as a linear imaging technique. The individual fiber-coupled lasers are then combined in an all-fiber coupler with an FC/PC connector. Each source is paired with a set of filters optimized for popular fluorophores that can be mounted in the PMT detection module.

The galvo-galvo scanner supports user-drawn scan geometries (lines, polylines, squares, and rectangles), custom photoactivation patterns (circles, ellipses, polygons, and points), and consistent pixel dwell times for better signal integration and image uniformity. Our intuitive interface …

The kit can be easily upgraded by adding a second PMT to the included dual-path epi-detection module with 8° collimation optics.

In confocal LSM, out-of-focus light is rejected through the use of a pinhole aperture, thereby leading to higher resolution. As shown in the drawing to the right, light enters the scan head through one of the laser inputs, is reflected onto the galvo-resonant scanner mirrors by the primary filter block, and exits the assembly through the wide field of view scan lens. Optical sections are formed by discretely measuring the signal generated within a specific focal plane. The laser source can also be controlled remotely by way of an auxiliary input.The detection module can be configured with our standard sensitivity multialkali PMTs or high sensitivity, ultra-low-noise GaAsP PMTs.

Thorlabs' MM201 Multiphoton Microscope Kit includes a galvo-resonant scanner, AR-coated scan lens for the visible and NIR wavelength ranges, epi-detection module, and a single GaAsP PMT.

Therefore, for a 1024 x 1024 pixel capture resolution, the scan field on the specimen would be ~111 μm x 111 μm. We also include a comprehensive installation and operation manual with basic preventative maintenance instructions to ensure that your system performs optimally for years to come. These CLS upgrades offer turnkey integration with virtually any inverted widefield microscope (not included) using a C-Mount camera port to access the intermediate image plane. Thorlabs' Modular Do-It-Yourself (DIY) ... Hardware Kit with All Components Required for Mounting to an Optical Table Ten 1/4"-20 and Twelve M6 Socket Head Cap Screws; Four 1/4" (M6) Washers; 2 mm, 2.5 mm, 3 mm, 4 mm, 5 mm, and 3/16" Hex L-Keys ; Four 2 mm (5/64") Hex Key Thumbscrews; MM201 Multiphoton Microscope Kit with Galvo-Resonant Scan Path. Alternatively, the 8 kHz galvo-resonant scanner is ideal for high-speed imaging applications with a maximum frame rate of 400 fps, while still utilizing the entire field of view.Each microscope kit has one GaAsP PMT with high quantum efficiency in the visible spectrum, providing elevated sensitivity to incoming illumination compared to photocathodes composed of alkali metals.

A 200um or 400um core fiber will lower the collection efficiency and make alignment more tedious. In an aberration-free imaging system, obtained by using the highest quality optical elements, the size of this focus spot, assuming uniform illumination, is a function of excitation wavelength (λIn actuality, the beam isn't focused to a true point, but rather to a bullseye-like shape.

For the MM201 DIY Galvo-Resonant Microscope Kit, the typical lead time is approximately 3 weeks from when the order is placed to when the kit is shipped out of Thorlabs' Newton, NJ Warehouse.These Multiphoton Microscope Kits can be controlled using the same software as our BergamoOptical Sections (Visualization of Thin Planes within a Bulk Sample)DIY Multiphoton Microscope Kit, Galvo-Galvo ScannerDIY Multiphoton Microscope Kit, Galvo-Resonant Scanner