The new expanded Chameleon portfolio gives you the ultimate choice of one-box solutions for brighter images, better sample viability and deeper tissue penetration covering the broadest range of flurophores. Choose the new Chameleon Vision with integrated precompensation or one of the Chameleon Ultra one-box lasers with the choice to add precompensation or a Chameleon OPO wavelength extension accessory later on. Whatever your choice, you will get the following Chameleon advantages to achieve brightest images in MPE.

Less brightness at 1000 nm at 9.4 mW	Increased brightness at 1030 nm at 9.4 mW	Highest brightness at longest wavelength - 1064 nm at 9.4 mW
Images courtesy of Jim Culver and Mary Dickinson, Baylor College

Brightest Images
Increasing the fluorescent signal is a key requirement for obtaining brighter images in MPE microscopy. The formula below shows two ways you can achieve brighter laser-excited images - either by increasing the average power or through decreasing the pulsewidth at the sample. More fluorescent signal means that more molecules in the sample are excited and this may decrease the sample viability through photo-toxicity. Even stained and fixed samples may photobleach when exposed to too much light for too long. Different samples behave in different ways. When linear damage (heating) is not important it is possible to increase the average power. When linear damage (heating) is dominant it may be more appropriate to decrease the pulse width at the sample and decrease the average power. With its high power and flexible pre-compensation, Chameleon gives you both choices.

Shortest Pulsewidths: Microscopes and scanning units may have highly dispersive elements like acousto-optics modulators (AOM) or complex, high-index lenses. It is now possible for some complex MPE optical systems to exhibit more than 30,000 fs² of GVD. This level of GVD will lengthen an unchirped pulse to many times the original duration by the time it reaches the sample. Using the formula above one can see that the longer pulses will decrease the fluorescent signal. This can be compensated by increasing the average power; however power on the sample in the order of higher than 100 mW may damage most samples. In these cases precompensating the pulses by adding a negative chirp is a good solution. Chameleon Vision enables you to manage your pulses and get the pulse duration that you think works best for your samples.

Longest wavelengths: Interest in using longer wavelengths has been increasing in the last few years. In addition to the availability of new fluorescent probes (like RFP, mCherry and mTomato), longer wavelengths are useful for SHG/THG microscopy. When used for standard MPE imaging they offer the advantage of increased flexibility of choices when you want to optimize fluorescent excitation, minimise damage to the sample or minimise light scattering in the sample. Chameleon Vision and Chameleon Ultra II tune up to 1080 nm to best reach the mFruit fluorescent proteins, while Chameleon OPO allows you to reach the limit of the water absorption around 1,400 to 1,500 nm and open new frontiers in SHG/THG and using newer fluorophores. The Chameleon platform gives you all the flexibility that you need now and in the future.

To learn how your experiment and imaging results could benefit from the industries broadest solutions portfolio for MPE, please contact our support team at 1-800-527-3786 or via email at tech.sales@coherent.com.

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