Confocal laser scanning microscopy (CLSM) is a widely used technique in the life sciences for making high-quality images of biological samples. The samples are illuminated by a focused laser beam which is scanned over the sample. in order to visualize fluorescent markers. Emission from each position is detected by a point detector and a pinhole removes out of focus emission light to improve contrast and resolution. CLSM results in optical sectioning and a 3D reconstruction of the sample is made.  

Two-photon microscopy is available on this instrument making use of a MaiTai Deep See laser unit. Two-photon microscopy is advantageous for deep imaging in thick (living) samples and is an alternative to confocal microscopy. In this type of microscopy, fluorophores are excited with light of a red shifted wavelength with half the energy needed for one photon excitation. The fluorophore needs to absorb multiple photons simultaneously tob e excitated. These events can only occur at high laser intensities which can be reached in the focal volume of a pulsed laser, thereby eliminating out-of-focus excitation and fluorescence. Two photon microscopy is advantageous for optical sectioning in thick samples.

Specifications:

· 4 lasers (405 nm, 488 nm, 561 nm, 640 nm)

· Galvos- and resonant- scanner

· Perfect Focus System

· 4 detection channels (2 GaASP and 2PMT detectors)

· 32-channel spectral detector unit

· Incubator with temperature regulation at the microscope level, CO2 and humidity at the cell culture level

· Multi-photon laser (Mai Tai TI:SAPPHIRE laser, 690-1040 nm)

· Non-descanned detectors

· Objective lenses:

o 60X water with automated correction ring (SR plan apo IR 60X WI, NA 1.3, WD 180µm)

o 60X oil (plan apo λ 60X oil, NA 1.4, WD 130 µm)

o 40X air (plan apo λ 40X, NA 0.9, WD 250 µm)

o 20X air (plan apo VC 20X, NA 0.8, WD 1000 µm)