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Combination of Optical Microscopy and AFM-
Collaboration Between PicoQuant and Bruker
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PicoQuant has combined its time-resolved confocal fluorescence microscope MicroTime200 with Brukers BioScope Catalyst Atomic Force Microscope (AFM). The synchronized acquisition of these two systems enables simultaneous recordings of AFM and optical images of the same sample region and makes new investigation schemes in the field of live cell imaging feasible.
Live Cell Imaging Feasibilities
The combined setup of the MicroTime 200 and the BioScope Catalyst is straightforward without the need of larger modifications of the two systems. The synchronized data acquisition enables scientists to analyze e.g. the impact of protein changes on cell shape and structure. It also allows high-resolution imaging by merging of sub nanometer topography with optically encoded functionality as well as investigations of inter- and intramolecular distances using force spectroscopy.
Technical Background
Brukers BioScope Catalyst AFM including its sample stage is mounted onto the inverse microscope body of PicoQuants MicroTime 200, which is configured for objective scanning. In this way, precise overlay of the confocal volume and the AFM tip can be realized. Electronic communication between the sample-scanner of the AFM and the data acquisition electronics of the MicroTime 200 enables simultaneous recordings with the two instruments. An optical alignment protocol has been developed and demonstrated by the synchronized acquisition of AFM and FLIM images of fluorescence TetraSpeck beads (drysurface), fixed glioblastoma cells (in liquids) as well as living HaCaT-cells (in liquids).
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Keywords: Atomic force microscopy BioScope Bruker FLIM Force Spectroscopy Life Cell Imaging Light Microscopy MicroTime 200 Optical Images PicoQuant Spectroscopy
PicoQuant GmbH
Rudower Chaussee 29/IGZ
12489 Berlin, Berlin
Germany
Tel: +49 30 6392-6568
Fax: +49 30 6392-6561
Web: http://www.picoquant.com
Imaging & Microscopy Issue 4 , 2012 as free epaper or pdf download
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