Setup of a fluorescence lifetime and spectral correlated acquisition system for two-photon microscopy

Spriet, Corentin; Trinel, Dave; Laffray, Sophie; Landry, Marc; Vandenbunder, Bernard; Heliot, Laurent; Barbillat, Jacques
December 2006
Review of Scientific Instruments;Dec2006, Vol. 77 Issue 12, p123702
Academic Journal
In this article we present a complete laser scanning microscope designed for simultaneous spectral and lifetime measurements from every point of the specimen located within the field of view. The pulsed laser source used for two-photon excitation provides good spatial resolution with minimal invasivity. In addition, the detection module was optimized for minimal photon loss, allowing laser power minimization and further reduction of cells photodamage. Analysis of biological samples illustrates the performances of this configuration, particularly when applied to fluorescent resonance energy transfer (FRET) measurements. Indeed, multiparametric acquisition is particularly useful to discriminate between FRET and artifactual response due to acquisition invasivity or cell heterogeneity. Combined with adapted homemade driving software, this system is stable, portable, and optimized for living cell studies.


Related Articles

  • Journal club. Wang, Michelle D. // Nature;8/21/2008, Vol. 454 Issue 7207, p921 

    The article discusses how light energy can be used as traps for microscopic particles. This light energy, according to the author, is the same energy that pushes dust particles away from the surface of the sun. Conventional optical traps, moreover, are made using laser beam and microscope....

  • TCSPC upgrade of a confocal FCS microscope. Benda, Aleš Hof, Martin; Wahl, Michael; Patting, Matthias; Erdmann, Rainer; Kapusta, Peter // Review of Scientific Instruments;Mar2005, Vol. 76 Issue 3, p033106 

    We extended the measurement capabilities of the Carl Zeiss ConfoCor 1 FCS microscope by (a) using pulsed picosecond diode lasers instead of a continuous wave (CW) laser excitation, (b) introducing a fast single photon avalanche diode detector, and (c) exploiting the capabilities of the PicoQuant...

  • A multiphoton laser scanning microscope setup for transcranial in vivo brain imaging on mice. Nase, Gabriele; Helm, P. Johannes; Reppen, Trond; Ottersen, Ole Petter // Review of Scientific Instruments;Dec2005, Vol. 76 Issue 12, p123702 

    We describe a multiphoton laser scanning microscope setup for transcranial in vivo brain imaging in mice. The modular system is based on a modified industrial standard Confocal Scanning Laser Microscope (CSLM) and is assembled mainly from commercially available components. A special...

  • Development of a periscopic confocal microscope for in situ observation under high magnetic fields. Hirota, Noriyuki; Ode, Takahiro // Review of Scientific Instruments;Mar2006, Vol. 77 Issue 3, p036107 

    This article contains a description of a periscope system developed for in situ high-resolution observation under high magnetic fields in combination with a confocal scanning laser microscope. It fits a typical cryocooler-operated superconducting magnet with a 100 mm diam room-temperature bore....

  • Automated Identification of Diatoms. Cairns, Jr., John; Almeida, Silverio P.; Fujii, Hitoshi // BioScience;Feb1982, Vol. 32 Issue 2, p98 

    Instrumentation permitting rapid automated identification of diatoms to be used in an environmental quality control system is described. A micro-optical processor, using input from an interference contrast microscope coherently illuminated by a Helium-Neon laser or an Argon laser, is the basic...

  • Microsurgery of the larynx. Sawyer, R. // Journal of the Royal Society of Medicine;Jul1980, Vol. 73 Issue 7, p473 

    The author reflects on the technological advancements in the diagnosis and treatment of diseases of the larynx with emphasis on the development of the laryngoscope. He discusses the importance of the microscope technique as a teaching tool through the use of a beam splitter and side arm viewing....

  • The 2010-2015 Outlook for Microscopes in India. Parker, Philip M. // Country Outlook Reports;1/ 2/2010, pN.PAG 

    Several charts on the 2010-2015 outlook for microscopes in several cities and states of India, including Delhi, Orissa and Assam, are presented.

  • Worlds That Might Have Been. Aronson, Stanley M. // Medicine & Health Rhode Island;Feb2010, Vol. 93 Issue 2, p39 

    In this article the author discusses the significant role of microscopes in developing medical science. He explains the importance of microscopes and other technical instruments in determining infective substances that affected the people during the medieval times. He argues that the instruments...

  • How to Avoid Errors in a Stereo Microscope Measurement System. Calahan, Tom // R&D Magazine;Jul2006, Vol. 48 Issue 7, p32 

    The article describes two potential which can arise when working with a stereo microscope, both leading to incorrect scaling of magnification. One is instrument reproducibility of the zoom setting and the other is human error where magnification setting is not properly transferred to the imaging...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics