BioScience AFM Acknowledgements
We would like to thank Josef Madl and Winfried Römer (BIOSS, Freiburg, Germany) for kindly providing the HeLa samples used for Figure 3 . The bacteriorhodopsin sample used for Figures 4 b, 5 a–c, 9 a, and 10 c–e was contributed by the group of Daniel J. Müller (ETH Zürich, BSSE Basel, Switzerland). The D96N mutant sample was supplied by Patrick Bosshart (Basel, Switzerland). The living cells used for measurements in Figures 7 , 8 , and 10 a–b were generously provided by the group of Andreas Herrmann (Humboldt University, Berlin, Germany). The authors would like to thank Tanja Neumann and Florian Kumpfe (JPK Instruments), as well as Jezz Leckenby (Talking Science) for the critical review during the preparation of the manuscript. Further assistance from Gabriela Bagordo (JPK Instruments) with the graphic design of the figures is gratefully acknowledged.
References [1] DJ Muller , Biochemistry 47 ( 2008 ) 7986 – 98 .
[2] A Berquand et al ., Microscopy Today 18 ( 2010 ) 8 – 14 .
[3] R Tomer et al ., Nat Methods 9 ( 2012 ) 755 – 63 .
Figure 10 : Quantitative Imaging™ of biological samples. Recording a complete force curve at each pixel with a pre-set vertical force eliminates lateral tip-sample forces. (a) An example with a very soft COS7 cell line with a low expression level of cytoskeletal fi laments. The complete set of recorded force curves unlocks a number of possibilities for offl ine analysis as demonstrated by the set-point height/topography (a) and sample indentation (b) at different forces. The online reconstruction of the set-point height at different forces allows either optimal resolution or nano-deformation of the sample, shown here with bacteriorhodopsin in buffer (c–e). Z -ranges in (a), (b), and (c–e) are 0–800 nm, 400–0 nm, and 0–600 pm, respectively.
SPM instruments today offer full integration with inverted light optical microscopes, giving researchers the flexibility of utilizing the benefits of both techniques in simultaneously acquired datasets. The integration with super-resolution microscopy pushes the boundaries even further by merging AFM and light optical microscopy into a seamless measuring system. The availability of specialized imaging modes helps to unravel structural problems that were not addressable by the AFMs of a decade ago. Finally, the emergence of fast-scanning AFM in combination with inverted light microscopy in a form that is directly applicable to life science research helps to get a more detailed understanding of dynamic processes in biology.
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[4] T Ando et al ., Proc Natl Acad Sci USA 98 ( 2001 ) 12468 – 72 .
[5] LM Picco et al ., Nanotechnology 18 ( 2006 ) 044030 .
[6] MB Viani et al ., Nat Struct Biol 7 ( 2000 ) 644 – 47 .
[7] T Ando , Nanotechnology 23 ( 2012 ) 062001 .
[8] DR Stamov et al ., Ultramicroscopy 149 ( 2015 ) 86 – 94 . [9] A Monserrate et al ., ChemPhysChem 15 ( 2014 ) 647 – 50 . [10] L Bozec et al ., Biophys J 92 ( 2007 ) 70 – 75 . [11] P Rahe et al ., J Vac Sci Technol B 28 ( 2010 ) C4E31 . [12] A Pyne et al ., Small 10 ( 2014 ) 3257 – 61 . [13] DJ Müller et al ., Biophys J 68 ( 1995 ) 1681 – 86 . [14] T Ando et al ., Chem Rev 114 ( 2014 ) 3120 – 88 . [15] HJ Butt et al ., EMBO J 8 ( 1989 ) 1657 – 63 . [16] J Klingauf et al ., Nature 394 ( 1998 ) 581 – 85 . [17] SW Schneider et al ., Proc Natl Acad Sci USA 94 ( 1997 ) 316 – 21 .
[18] T Fukuma et al ., Rev Sci Instrum 77 ( 2006 ) 123703 .
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