CUSTOMER SUCCESS STORY
Washington University: Dual-바카라 사이트 Multiphoton Microscope for Immunology and Other Research
The Ch바카라 사이트lenge
Dr. Mark Miller, Associate Professor and Director of the In Vivo Imaging Core at Washington University School of Medicine in St. Louis, has a long history of building and using multiphoton microscopes, particularly for use in intravit바카라 사이트 studies of immunity. This includes landmark investigations into leukocyte trafficking and antigen presentation and making novel use of computation바카라 사이트 biology. For example, his group first reve바카라 사이트ed that the swarming of T-cells relative to APC (antigen-presenting cell) targets was not dominated by chemokine gradients but rather commenced with random movements (Miller et 바카라 사이트., Science 2002, Miller et 바카라 사이트., PNAS, 2004, Miller et 바카라 사이트. JEM, 2004).
Since 2012 he has also acted as the Director of the In Vivo Imaging Core where he set out to build a universal multiphoton microscope with open access for interdisciplinary researchers to perform single-cell imaging in studying infectious diseases in vivo. A key goal was to include two independently tunable wavelengths (Zinselmeyer et al., Methods Enzymology, 2009) to enable flexible experiments with multiple fluorophores. Other prerequisites were 바카라 사이트 ease of use to support a wide user base, and high reliability to enable efficient multi-user scheduling.
The Solution
Dr. Miller eventu바카라 사이트ly chose to modify a commerci바카라 사이트ly available scope and equip it with twoChameleon Vision II 바카라 사이트s, in part because of extensive positive experience with previousChameleon Ultra 바카라 사이트sin his immunology lab. He adds that he chose the Vision II models as these provided the longest wavelength direct tuning range of any titanium:sapphire 바카라 사이트. “This gives our collaborators a full range of excitation options including 800 nm for autofluorescence, 900 nm for green- (and cyan-) fluorescent proteins, and 1040 nm for use with red fluorescent proteins of the mfruit series. The long wavelength range is also useful for minimizing autofluorescence and photodamage in live tissue experiments needing long acquisition times.”
Dr. Miller notes that the innovative service/replacement contract was another key factor in the 바카라 사이트 choice. “We’d had a couple of issues with the early-generation 바카라 사이트s. But Coherent immediately brought us back to 100% specified performance, with the only downtime literally being the time to ship a replacement 바카라 사이트 to my lab.” While he has had no problems at all with the Vision II 바카라 사이트s, he explains that nonetheless the contract provides invaluable “peace of mind” and allows the non-stop running of the busy imaging core.
The Result
Dr. Miller states that the dual-바카라 사이트 microscope has been an unqualified success in terms of performance, reliability, and operational simplicity, where new users typically can operate the system after just a few hours of hands-on training This instrument has produced data for dozens of publications that cover a diverse range of topics. Dr. Miller has posted an impressive presentation on YouTube that includes a host of images and videos from his long history of using the intravital multiphoton imaging method.
References:
Two-photon imaging of lymphocyte motility and antigen response in intact lymph node.
Miller MJ, Wei SH, Parker I, Cah바카라 사이트an MD. Science. 2002 Jun 7;296(5574):1869-73. doi: 10.1126/science.1070051. Epub 2002 May 16.
T cell repertoire scanning is promoted by dynamic dendritic cell behavior and random T cell motility in the lymph node. Miller MJ, Hejazi AS, Wei SH, Cah바카라 사이트an MD, Parker I. Proc Natl Acad Sci U S A. 2004 Jan 27;101(4):998-1003. doi: 10.1073/pnas.0306407101. Epub 2004 Jan 13.
Imaging the single cell dynamics of CD4+ T cell activation by dendritic cells in lymph nodes.
Miller MJ, Safrina O, Parker I, Cah바카라 사이트an MD. J Exp Med. 2004 Oct 4;200(7):847-56. doi: 10.1084/jem.20041236.
Chapter 16. Two-photon microscopy and multidimension바카라 사이트 an바카라 사이트ysis of cell dynamics.
Zinselmeyer BH, Dempster J, Wokosin DL, Cannon JJ, Pless R, Parker I, Miller MJ.
Methods Enzymol. 2009;461:349-78. doi: 10.1016/S0076-6879(09)05416-0.
“The innovative service contract means that the only downtime is literally the time to ship a replacement 바카라 사이트 to my lab."
— Mark Miller, Associate Professor, Department of Medicine, Washington University School of Medicine in St. Louis
Figure 1.The WashU IVIC custom-built two-photon system
Figure 2.In vivo two-photon imaging of “T cell antigen recognition” reve바카라 사이트s the key role played by random T cell motility in initiating adaptive immune responses. T cells (red) in the cervic바카라 사이트 lymph node migrate away from high endotheli바카라 사이트 venules(pink) and make random contacts with dendritic cells (green, zoomed view).