How a Multiphoton Microscope Works & How We Save You Time & Money
Even with the many equipment variations and types, Excedr’s leasing program is able to accommodate any preferences your lab may have with regards to brand or model. Get in touch with us today and learn about how leasing can discount your three-photon microscope’s price.
All equipment brands/models are available
The Advantages of Excedr’s Multiphoton Microscope Leasing Program:
- Eliminates the upfront cost of purchasing equipment by spreading its cost over time
- Minimizes equipment downtime with included complete repair coverage and preventive maintenance
- Takes advantage of potentially 100% tax deductible* payments, providing you significant cash-savings
- Expedites the administrative work needed for instrument procurement and logistics
- Conserves working capital, enabling you to reinvest in your core business and operations (staffing, inventory, marketing/sales, etc.)
- Accommodates all manufacturer and model preferences
*Please consult your tax advisor to determine the full tax implications of leasing equipment.
Microscopy is the study of small objects with the use of microscopes.
Most optical microscopes have the illumination source underneath the sample. The illumination sources are pointed at the objective lens which magnifies the image making it easier to see with the human eye. Other microscopy techniques exist, however, that use different light sources and a variety of configurations. For example, fluorescence microscopy employs fluorescent tagging to illuminate specific parts of the sample rather than having an external light source. The fluorescent tags are then “activated” by using electromagnetic radiation which illuminates the sample and allows for image detection and magnification to occur.
When imaging relatively thick samples with fluorescent microscopy, background fluorescent light can negatively affect the image. If you only want to observe something on the surface of a thick specimen, the background fluorescent light from the thickness of the material would drown out the fluorescent light from the surface of the material. Multiphoton microscopy, a nonlinear optical technique, solves this problem by using two photons, rather than the one which is standard in fluorescence microscopy, to excite the sample. By using two-photon beams, this technique can create detailed 3D images of living cells while avoiding photobleaching and phototoxicity. Because of its use of two-photon beams, this technique is also known as two-photon excitation microscopy or non-linear excitation microscopy.
In multiphoton microscopy, the sample is still dyed with fluorophores as is done in conventional fluorescent microscopy and then the two-photon beams are focused on a specific region of the object. The two photons that are used can be of the same or different frequencies but they must strike the molecule within one femtosecond of each other. Due to the precision needed in this last requirement the focused laser that is used must be able to function at very high pulse rates. These lasers are often called femtosecond lasers. When the two photons do strike the molecule it excites that atom’s electrons to a higher, less stable state. This energy difference between the two states is equal to the sum of the energies of the two photons that were used to excite it. This means that only the specific area that is struck by both photons will be excited and fluoresce. This localized excitation and fluorescence is one of the reasons that multiphoton excitation microscopy is such a powerful tool. The other main benefit of using two-photon excitation microscopy is that it allows for optical sectioning at greater depths. It is able to do this due to the extreme localization of the lasers used which allows for greater depth penetration. Additionally, the light used is usually red and infrared which exhibits less scattering than blue or bluer light.
Multiphoton Microscopy Variations, Techniques, & Price
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To further reduce the out of focus fluorescent light and decrease the amount of tissue scattering, three-photon excitation microscopy can be employed. Three-photon and two-photon excitation microscopy have very similar procedures. The obvious major difference is that three photons must be absorbed by the molecule before it fluoresces rather than just two. This also means that the wavelengths of the photons used in three-photon microscopy are much longer (about 1300 nanometers) than those used in two-photon excitation microscopy (about 910 nanometers). Four-photon excitation is currently being looked into as well, however, it has not found any major practical biological use yet.
Second-Harmonic Imaging (SHIM)
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Whenever fluorophores and fluorescence are used to image an object, phototoxicity and photobleaching become issues that need to be taken into account. Second-harmonic imaging microscopy is a nonlinear optical technique that exploits the second-harmonic generation effect. This means that no molecules are excited and so phototoxicity and photobleaching are nonissues. SHIM is able to look at cells and living tissue’s structure by creating contrast by looking at variations in a material’s ability to create second-harmonic light. Second-harmonic light occurs when two photons, of the same frequency, strike a material, combine, and generate a new photon of twice the energy of the initial photons. SHIM can be used to create 3D images of samples even if they are relatively thick. Additionally, because organic structures produce second-harmonic generation signals naturally, tagging is often not required. This means that the sample can be observed unaltered.
Maria Goeppet-Mayer: Two-Photon Absorption
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Two-photon excitation microscopy is based on the reaction of two-photon absorption. This was first predicted in 1931 by Dr. Maria Goeppert-Mayer while at the University of Göttingen in her doctoral thesis. It would, however, take thirty more years and the invention of the laser before her theory could be verified. On her doctoral committee, there were three Nobel Prize winners: her advisor Max Born, James Franck, and Adolf Otto Reinhold Windaus. In recognition of her work, the unit for a two-photon absorption cross-section is named after her, the Goeppert Mayer. She would continue her scientific career in the United States as a teacher at Sarah Lawrence College in the 1940s. While there, she joined the Manhattan project and worked with yet another Nobel Prize winner Harold Urey. After the war ended she found herself in Chicago as an associate professor in the Physics Department of the Institute for Nuclear Studies. She was recognized for her intellect and eventually was offered another position at the Argonne National Laboratory to which she replied, “I don’t know anything about nuclear physics”. It was during her time in Chicago where she penned her theory on the nuclear shell model. In it, she mathematically explained why certain configurations in an atom’s nuclei result in a stable configuration. For this she would later be awarded the Nobel Prize in physics, becoming the second woman to do so after Marie Curie.
Your lab’s ability to function should not be limited by how expensive its equipment is. Whatever your leasing needs, be they financing a C-arm or multiphoton microscope, we’re here to help. We can discuss your financing options in further detail. Call us at (510) 982-6552 or fill out our contact form online.
We Offer Non-Linear and Two-Photon Excitation Microscope Leases to Fit Every Need
This off-balance sheet financing structure provides three options at the end of the term. The lessee has the option to return the equipment to the lessor, renew at a discounted rate, or purchase the instrument for the fair market value. Monthly payments are also 100% tax deductible which yields additional monetary savings.
If you recently bought equipment, Excedr can offer you cash for your device and convert your purchase into a long-term rental. This is called a sale leaseback. If you’ve paid for equipment within the last ninety days, we can help you recoup your investment and allow you to make low monthly payments. This also frees up money in your budget rather than tying it down to a fixed asset.
Two-Photon Excitation Microscope Manufacturers & Models on the Market
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A1R MP+, A1 MP+
Ultima series, Ultima 2Pplus, Ultima In Vitro, Ultima Investigator
Movable Objective Microscope (MOM), 3P-MOM, DF-SCOPE