Maitai hp laser

Manufactured by Spectra-Physics

Sourced in Japan, Canada

The MaiTai HP laser is a mode-locked ultrafast laser system designed for laboratory applications. It generates ultrashort pulses at a wavelength of 800 nm with a typical pulse duration of less than 100 femtoseconds. The laser features high average power output and a repetition rate of 80 MHz, making it suitable for various experimental setups requiring ultrafast laser sources.

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Lab products found in correlation

Lsm 710, by Zeiss (1 mentions) Sp5 sted microscope, by Leica (1 mentions) Atto 647n, by Active Motif (1 mentions) Lsm 880, by Zeiss (1 mentions) Laurdan, by Thermo Fisher Scientific (1 mentions) Tcs sp5 confocal microscope, by Leica (1 mentions) Laurdan, by Spectra-Physics (1 mentions) 0.8 na objective, by Nikon (1 mentions) Fluorescein conjugated dextran, by Merck Group (1 mentions) Movable objective microscope, by Sutter Instruments (1 mentions)

Close spelling variants of this product

MaiTai (104 citations) Titanium: sapphire femtosecond MaiTai HP laser (2 citations) MaiTai HP IR laser (2 citations)

7 protocols using maitai hp laser

Imaging Axons in Frontal Cortex

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To image descending frontal axons, we used an Ultima IV laser scanning microscope (Bruker, Middleton, WI) and a 40 × 0.8 NA water immersion objective (Olympus, Center Valley, PA). A Mai Tai HP laser (Spectra physics, Santa Clara, CA) was tuned to 910 nm in order to excite GFP. We imaged axon segments (∼40 microns in length) and obtained image stacks with a 1 μm z-step. To image ascending BLA and OFC axons that overlap in the L2/3 region of the dorsal frontal cortex (sampling regions labeled with GFP and td-tomato in the same mouse), we used a Zeiss LSM 710 laser scanning confocal microscope using 40 × 0.8 NA oil immersion objective. Axon length sampling was comparable across MS and control groups (Supplementary Table 1).

Thomas A.W., Delevich K., Chang I, & Wilbrecht L. (2019). Variation in early life maternal care predicts later long range frontal cortex synapse development in mice. Developmental Cognitive Neuroscience, 41, 100737.

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STED Microscopy Protocol and Setup

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For stimulated emission depletion (STED) microscopy and the corresponding confocal microscopy a LEICA SP5-STED microscope was used. The fluorophores ATTO647N (Activemotif) or MegaRed (Sigma) were excited with an 80 MHz pulsed diode laser at 635 nm or 531 nm respectively (Pico-Quant), (pulse width < 100 ps). STED depletion was achieved using a mode-locked titanium sapphire laser (Spectra Physics Mai Tai HP laser) operating at 770 nm with a repetition rate of 80 MHz. The delay between the excitation and STED pulses was adjusted electronically to optimize depletion. The excitation and the STED beams were focused by a 100× oil immersion objective (NA 1.4 PL APO STED, 100×; Leica Microsystems). The fluorescence signal was collected by the same objective and detected confocally between 650 and 690 nm for ATTO647N and between 540 and 595 nm for Megared with an APD detector. Using this set up, a resolution of ˜250 nm in the confocal images and 50–70 nm in the STED images was achieved. Except for contrast stretching and level adjustment no further image processing was applied.

Shi C.S., Qi H.Y., Boularan C., Huang N.N., Abu-Asab M., Shelhamer J.H, & Kehrl J.H. (2014). SARS-CoV ORF-9b suppresses innate immunity by targeting mitochondria and the MAVS/TRAF3/TRAF6 signalosome. Journal of immunology (Baltimore, Md. : 1950), 193(6), 3080-3089.

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Two-Photon Lifetime Imaging of Coumarin 6

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Fluorescence lifetime images were collected with a Zeiss LSM 880 (Carl Zeiss Microscopy) microscope equipped with a Spectra-Physics MaiTai HP laser (Spectra-Physics) for two-photon excitation, tuned at 800 nm, and collected using a Zeiss 63×/1.41 NA oil objective. The fluorescence signal was collected by a photomultiplier tube (H7422P-40, Hamamatsu) and recorded using a FLIMbox (model A320 ISS) to obtain the lifetime information. The pixel dwell time for the acquisitions was 16 μs and the images were taken with sizes of 256 × 256 pixels, accumulating 15 frames. The data from each pixel were recorded and analysed using the SimFCS software (Laboratory for Fluorescence Dynamics, University of California, Irvine, CA) to perform the phasor transformation and with custom MATLAB code to quantify the lifetime. The phasor position was corrected for the instrument response function by acquiring the fluorescence of a solution with a known lifetime (0.4 mM Coumarin 6 dissolved in ethanol, τ = 2.5 ns). The lifetime was determined as τ = s/ωg, where τ is the fluorescence lifetime, g and s are the phasor coordinates and ω is the laser frequency (80 MHz).

Agustinus A.S., Al-Rawi D., Dameracharla B., Raviram R., Jones B.S., Stransky S., Scipioni L., Luebeck J., Di Bona M., Norkunaite D., Myers R.M., Duran M., Choi S., Weigelt B., Yomtoubian S., McPherson A., Toufektchan E., Keuper K., Mischel P.S., Mittal V., Shah S.P., Maciejowski J., Storchova Z., Gratton E., Ly P., Landau D., Bakhoum M.F., Koche R.P., Sidoli S., Bafna V., David Y, & Bakhoum S.F. (2023). Epigenetic dysregulation from chromosomal transit in micronuclei. Nature, 619(7968), 176-183.

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Laurdan-based Membrane Fluidity Imaging

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Cells labeled with Laurdan (10 μM, Molecular Probes) were visualized with Leica TCS SP5 confocal microscope. Submembraneous areas were defined as areas extending 1 μm towards the cell interior. Laurdan was excited at 800 nm with a mode-locked titanium sapphire laser (MaiTai HP laser, Spectra-Physics). An amplitude modulator linked to the Leica Microsystems software system controlled two-photon intensity input. To record intensities at selective wavelengths, internal photon multiplier tubes collected images as 8-bit, unsigned images with 512 × 512 pixels at 400 Hz scan speed. Laurdan image calculations were carried out in ImageJ (NIH). The generalized polarization (GP) was defined as

and calculated for each pixel using the two Laurdan intensity images.

Pfeiler S., Khandagale A.B., Magenau A., Nichols M., Heijnen H.F., Rinninger F., Ziegler T., Seveau S., Schubert S., Zahler S., Verschoor A., Latz E., Massberg S., Gaus K, & Engelmann B. (2016). Distinct surveillance pathway for immunopathology during acute infection via autophagy and SR-BI. Scientific Reports, 6, 34440.

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Large Animal 2P Microscope Design

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The 2P microscope for imaging swine was designed with four key specifications: 1) ability to accommodate large animals with considerable flexibility, 2) intensity and fluorescence lifetime imaging without switching detection hardware and scan path, 3) externally triggered high speed volumetric scanning, and 4) low-cost construction from off-the-shelf parts. The overall design of the microscope was based heavily on the previously published open source TIMAHC (Two-photon Imaging that is Modular, Adaptable, High-performance and Cost-effective) design^{14 (link)} with exceptions detailed below. All 2P imaging was performed using excitation light from a MaiTai HP laser (Spectra Physics, Milpitas, CA) 16× , 0.8 N.A. objective (Nikon Instruments, Tokyo, Japan). Emission light was filtered through 440/80 (cyan fluorescent protein and the chloride-sensitive organic dye), 525/50 (YFP and GFP), or 605/70 (TurboRFP and tdTomato, Chroma Technology, Bellows Falls, VT).

Costine-Bartell B.A., Martinez-Ramirez L., Normoyle K., Stinson T., Staley K.J, & Lillis K.P. (2023). 2-Photon imaging of fluorescent proteins in living swine. Scientific Reports, 13, 14158.

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Chronic Two-Photon Imaging of Axons

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The procedure for imaging through a chronic cranial window using two-photon imaging was performed as previously described40 (link). Mice were anaesthetized with isoflurane anaesthesia, and a bar affixed to the skull was screwed into a metal post and fixed to a metal base. The brain was imaged using an Ultima IV laser scanning microscope (Prairie Technologies) and a water immersion 40x magnification 0.8 numerical aperture objective. A Mai Tai HP laser (Spectra Physics) was tuned to 910 nm for excitation of green fluorescent protein. Approximately, 80 μm segments of axon were imaged at zoom 4 with high resolution (12.05 pixels per μm) within 100 μm of the surface (layer 1). Image stacks were obtained using a 1-μm z-step. Imaging for the main experiments was within 0.8 mm of the midline in the right hemisphere, contralateral to the viral injection. A smaller set of regions of interest were studied in the ipsilateral left hemisphere (0–0.8 mm from midline), and lateral right hemisphere (0.8–1.4 mm) for comparison of density and turnover (Supplementary Fig. 2). For relocation of the same axons across imaging sessions, a bright-field image and a two-photon image stack were taken of the pattern of the blood vessels and neuronal processes as a reference point. After imaging, mice were given subcutaneous saline and allowed to recover in a separate cage before returning to the homecage.

Johnson C.M., Peckler H., Tai L.H, & Wilbrecht L. (2016). Rule learning enhances structural plasticity of long-range axons in frontal cortex. Nature Communications, 7, 10785.

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Two-Photon Microscopy of Brain Vasculature

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Animals were imaged using a two-photon microscope consisting of a Movable Objective Microscope (Sutter Instruments, CA) and a MaiTai HP laser (Spectraphysics, Mountain View, CA), controlled by MPScan software (Nguyen et al. 2006 (link)). A 20× 0.5 N.A. (Olympus, Center Valley, PA), or 20 × 1.0 N.A. (Olympus) water dipping objective was used for imaging. Before each imaging session, animals were briefly anesthetized with isoflurane and were infraorbitally injected with 50µL (50mg/mL) fluorescein-conjugated dextran (70 kDa; Sigma, St. Louis, MO) or rhodamineB-conjugated dextran (70 kDa; Sigma). The laser was tuned to 800nm for imaging fluorescein alone, and 910nm for rhodamineB/GCaMP3 imaging. For isoflurane vasodilation experiments, mice were placed on a homoeothermic heating pad while anesthetized with 2% isoflurane in air. Imaging sessions typically lasted ~2 hours. Each vessel was imaged for approximately 15 minutes at ~8 frames/second. Penetrating vessels were imaged 30–250 µm below the pia. We were able to image capillaries clearly down to 200µm through PoRTS windows with no loss of resolution (Supplementary Fig. 6; Supplementary Table 1). Arterioles and venules were identified morphologically (Blinder et al. 2010 ).

Gao Y.R., Greene S.E, & Drew P.J. (2015). Mechanical restriction of intracortical vessel dilation by brain tissue sculpts the hemodynamic response. NeuroImage, 115, 162-176.

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