2-photon microscopy in Neuroscience
2-photon fluorescence microscopy has become a key technology in biological imaging enabling three-dimensional, noninvasive studies of biological tissue on the submicron scale. The contrast mechanism in 2-photon microscopy is based on the excitation of fluorophores by two photons, typically in the infrared spectral range. Upon excitation the fluorophores relax back by emitting a photon in the visible which is detected.
High peak power of the laser is key to drive this nonlinear process. Therefore femtosecond lasers with clean temporal pulse shape and average output powers of >1W are essential for such applications. In addition integrated power control and dispersion precompensation are a must to enable maximum image brightness in the experiment. To get a better idea of the requirements on two-photon microscopy we recommend the following white paper and webinar.
In contrast to conventional linear fluorescence microscopy, the nonlinear character and the longer excitation wavelength in multi-photon microscopy offer several advantages: (i) larger probing depth (ii) lower phototoxicity and reduced photodamage. Both advantages make 2p-fluorescence microscopy an ideal choice for the direct spatiotemporal visualization of neurons and neuronal activity in living animals. The key fluorescent proteins for such experiments are green and red fluorescent proteins that can be excited by 2-photon absorption at 920nm and 1050nm, respectively. The clear requirements in terms of wavelength, pulse duration, and output power make TOPTICA's easy-to-use and fully integrated fiber lasers FemtoFiber ultra 920 at 920nm and FemtoFiber ultra 1050 at 1050nm the perfect choice for 2-photon experiments in neuroscience.
To understand the potential and latest trends in 2-photon fluorescence microscopy in neuroscience please watch the webinar with Dr. Weijian Zong from the Moser lab in Trondheim, pioneering the field of miniaturized 2-photon microscopy:
WEBINAR:
MICROSCOPY IN NEUROSCIENCE
THU NOV 11, 6:30 PM - 7:30 PM GMT +1 (1 HOUR)
2-photon Optogenetics in Neuroscience
All-optical interrogation is a novel approach to understand how active patterns in neuronal activity drive behavior. In such experiments the visualization of neuronal activity by 2-photon imaging is combined with 2-photon optogenetics to stimulate individual neurons by photoactivation of channelrhodopsins within the cell.
With the need of exciting many neurons in parallel, the laser requirements and microscopy technology for 2-photon optogenetics are fundamentally different from 2-photon imaging. Typically, high-power multi-Watt lasers at 1030-1040nm with repetition rates in the 100kHz - 1MHz range are used in combination with a spatial light modulator (SLM) to excite 10s to 100s of neurons simultaneously. Such lasers ideally offer integrated power control and dispersion precompensation to optimize 2-photon excitation efficiency.
TOPTICAs FemtoFiber vario 1030 HP has been fully optimized to perfectly match these requirements and, together with the FemtoFiber ultra 920, complements TOPTICA's product offering for all-optical interrogation experiments in neuroscience.
Thu Nov 11, 6:30 PM - 7:30 PM GMT +1 /
5:30 AM- 6:30 AM Melbourne, Sydney 5:00AM - 6:00AM Adelaide (1 Hour)
Microscopy in Neuroscience
Part I Dr. Zong
High-resolution calcium imaging with 2P miniscopes in freely-moving mice
Due to the unsuitability of benchtop imaging for tasks that require unrestrained movement, investigators have tried, for almost two decades, to develop miniature 2P microscopes-2P miniscopes–that can be carried on the head of freely moving animals. In this talk, I would first briefly review the development history of this technique, and then report our latest progress on developing the new generation of 2P miniscopes, with resolution, speed, and z-scanning capability comparable to that of 2P benchtop microscopes. The new technique permits large-scale and high-resolution calcium imaging in freely-moving mice, and opens the door to investigate brain functions during animal`s most natural behaviors. Current limitations and future developments will also be discussed.
Part II Dr. Eisele
Easy-to-use fiber lasers for 2-photon microscopy in neuroscience
In this Webinar Dr. Max Eisele will give a quick overview on TOPTICAs laser technology for fluorescence and 2-photon fluorescence microscopy. This general introduction is followed by a detailed discussion on the key requirements for excitation sources used in two-photon fluorescence microscopy and two-photon optogenetics in neuroscience performing all-optical interrogation of neuron interaction. The presentation will be finalized by introducing TOPTICAs laser for neuroscience that offer a complete solution for researchers working in the field of 2-photon microscopy and 2-photon optogenetics. These lasers offer highest performance while significantly simplifying the laser operation and the entire experimental setup. Don’t miss this opportunity to learn about TOPTICAs lasers for neuroscience and how they simplify your research!
Who Should Attend This Webinar?
- Scientists and companies working in the field of 2-photon fluorescence microscopy and neuroscience
- Scientists working in the field of 2-photon optogenetics and neuroscience
- Scientist and companies who are looking for easy-to-use and fully optimized laser engines for their research or product lines
In This Webinar You Learn -
- Learn about the latest trends using miniaturize 2-photon microscopes in neuroscience
- Learn about essential laser requirements for 2-photon fluorescence excitation and 2-photon optogenetics in neuroscience
- Learn about the ideal lasers for all-optical interrogation in neuroscience
- Learn about easy-to-use laser engines optimized for two-photon fluorescence excitation and 2-photon optogenetics.
Further information:
TOPTICA: FemtoFiber ultra 920 Femtosecond fiber laser for 2-photon microscopy and neuroscience
TOPTICA: Pulse on Demand: FemtoFiber vario 1030